Abstracts of Oral Communications and Posters International ... · PDF file10th International...

R E V I S T A P O R T U G U E S A D E P N E U M O L O G I A

Vol XIII Suplemento 2 Outubro 2007

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10th International Conference on Brochoalveolar Lavage// Abstracts of Oral Communications and Posters

10th International Conference on Bronchoalveolar Lavage

June 17-19, 2006Coimbra, Quinta das Lágrimas

Abstracts of OralCommunications (C)

Abstracts of Oral Communications and Posters

C01Identification of bio-markers insarcoidosis bal using multidimensionaldifferential display proteome analysiscoupled with nano-hplc ms/msOtto F Villa1, Oscar E Guevara1, Marc Judson2,

Robert P Baughman3, and Michael Iannuzzi 1

1 Mount Sinai School Of Medicine. New York, NY;2 Medical University of South Carolina. Charleston, SC.3 University of Cincinnati Medical Center, Cincinnati, OH,

United [email protected]

AbstractIt is well recognized that activated T-cells together with his-tiocyte/macrohages accumulate in the interstitium and al-veoli of the sarcoidosis lung forming granulomas. The goalof this study is to perform a global analysis of the sarcoido-sis-macrophage proteome using multidimensional differen-tial display to identify molecular species unique to this ILD.To this effect CD14+ cells were isolated from the BAL ofsarcoidosis patients (n=10), patients with other ILDs (n=10)or from patients with no ILD (n=4). Proteomes of eachBAL were prepared using large format two-dimensional gelelectrophoreses. TiFF files were warped in reference to sar-coidosis and all spots integrated into a large matrix.A total of 23960 spots were analyzed, 1198 spots compri-sed the sarcoidosis proteome, of these 383 were up-regula-ted over two-fold and 294 over 10 fold when compared withproteomes of the patients without ILD (Fig. 1). The sarcoi-dosis proteome was next compared with all other ILDs in astep-wise manner or globally, showing that 154 sarcoidosisspots remained upregulated over 3 fold and 94 over 10 fold.Protein identification by nano-HPLC-MS/MS on an LTQinstrument revealed species involved in cellular metabolism,

calcium mobilization, actin binding, cell signaling, apopto-sis, cell division and inflammation of which 10 are beingstudied as candidate biomakerkers.

C02The clinical significance of IL2, IL12,TGFbeta serum and balf levels insarcoidosis-a preliminary studyA Goljan-Geremek1, A Skoczylas1, U Demkow 2,

E Puscinska1, M Polubiec1, I Bestry1, D Gorecka1

1 Dept. of Respiratory Medicine, Institute of TB and Lung Di-seases, Plocka, Warsaw, Poland

2 Dept. of Laboratory Diagnostics and Clinical Immunology,Warsaw Medical University, [email protected]

AbstractThere is evidence from clinical immunology that the serumand BALF cytokine analysis may be useful for clinicians toassess the extent and activity of sarcoid inflammation.Aim: To test the relationship between serum and BALF IL2,IL12, TGFbeta levels and the clinical outcome of sarcoidosis.Methods: In 38 patients with biopsy proven sarcoidosis wemeasured the serum and BALF IL2, IL12, TGFbeta levelsby the commercial sandwich ELISA kit from BioSource andthe non specific serum inflammation markers (proteinogram,CRP, D-dimer, Fibrinogen,)We compared the serum and BALF IL2, IL12, TGFbetalevels, serum non specific inflammation markers in sar-coid patients according to different clinical signatures ofthe disease.Results: The EN (n=5) was associated with high serumalfa2globulin (p=0,03) and fibrinogen level (p=0,0002) andlow 02saturation during 6mWT (p=0,04).



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C03Cytokine release from alveolarmacrophages in idiopathicinterstitial pneumoniaQiao Ye, Ulrich Costabel, Josune Guzman1

Department of Pneumology and Allergy, Ruhrlandklinik, Essen,Germany1 General and Experimental Pathology, Ruhr University, Bochum,

[email protected]

AbstractObject of study: Idiopathic interstitial pneumonia (IIP) is agroup of interstitial pneumonias with unknown causes, clas-sified as different subgroups according to the histological cha-racteristics. Alveolar macrophages (AM) are able to releasevarious cytokines and play an important role in the inflamma-tory and fibrotic process of IIP. However, the production ofcytokines by AMs in the subgroups of IIP is still unclear.Methods: We measured the release of TNF-alpha, TGF-beta,IL-1beta, IL-6, IL-10, IL-12 and IL-18 from bronchoalveolarlavage (BAL) macrophages in 9 patients with cryptogenic or-ganising pneumonia (COP), 13 with non-specific interstitialpneumonia (NSIP), 5 with respiratory bronchiolitis interstiti-al lung disease (RBILD), 4 with desquamative interstitial pneu-monia (DIP), 19 with idiopathic pulmonary fibrosis (IPF) and10 controls. AMs were cultured for 24h with RPMI mediumalone or with lipopolysacchride (LPS) (100ng/ml). Cytokinesin the supernatants were measured by ELISA.Results: The spontaneous levels of TNF-alpha, TGF-beta,IL-1beta, IL-6, IL-10, IL-12 and IL-18 released from BALmacrophages were significantly higher in COP than in IPFand controls (p<0.05 or <0.01 respectively), and except forIL-18 also significantly higher in NSIP than in IPF and con-trols (all p<0.05). The production of these cytokines withor without LPS stimulation was highest in COP, followedby NSIP, and lowest in DIP/RBILD and IPF.

C04Characteristics of myofibroblasts(MyFb) recovered by bronchoalveolarlavage in interstitial lung diseases (ILD)Fireman E

Department of Pulmonary and Allergic Diseases, 2NationalLaboratory Service of Interstitial Lung Diseases, Sackler Facultyof Medicine, Tel Aviv University, Tel Aviv, [email protected]

AbstractWe have developed a novel technique which allows to isola-te myofibroblasts (MyFb)cells from the fluid recovered du-

ring bronchoalveolar lavage in patients undergoing this pro-cedure for diagnostic porpoises. This method is crucial sin-ce it gives us the possibility to achieve those cells also fromentities (like sarcoidosis, hypersensitivity pneumonitis) wherethere is no clinical indication for open lung biopsy or thora-coscopy. Most of the studies in literature use animal model,cell lines or cells recovered from lung surgery. The pheno-type of MyFb in sarcoidosis (SA) and idiopathic pulmonaryfibrosis (IPF) is unclear. We characterized the cytoskeletalproteins and the contraction properties in alveolar-associa-ted fibroblasts recovered by bronchoalveolar lavage (BAL)in the two diseases. MyFb were studied from BAL cells ineight IPF and seven SA patients. Cytoskeletal proteins wereidentified by ELISA and immunofluorescent methods. Bio-chemical measurements were done by dry chemistry. Con-traction was performed by a gel contraction assay. MyFbalpha-SM actin measured by ELISA was higher in IPF thanin SA. Vimentin, desmin, myosin, and fibroblast markerswere expressed equally. Only in IPF did the MyFb reveal themyofibroblast phenotype showing alpha-SM actin immu-nofluorescence labeling and, by electron microscopy, fila-ments with associated dense bodies with rough endoplas-mic reticulum. Gel contraction showed that cells in IPFcontracted significantly more than in SA. This may explainthe difference in the behavior patterns and outcomes of thetwo diseases. Recovery of MyFb from BAL fluid open awindow to the mesenchymal cells research in ILD.

C05Pulmonary alveolar proteinosis.A report of turkish casesN Mogulkoc, O Ozdemir, E Eryuksel1, N Kokturk2,

M Yuksel1, S Karakurt1, T Celikel1

Department of Chest Medicine, Ege University, Izmir, Turkey;1 Marmara University Hospital, Istanbul;2 Gazi University Hospital, Ankara, [email protected]

AbstractWe review 15 patients (5 male and 10 female, mean age 37years) with the diagnosis of pulmonary alveolar proteino-sis diagnosed and followed in three University Hospitalsin Turkey. The major clinical features were dyspnea, cou-gh, and chest pain, with durations of symptoms from threemonths to five years. On chest radiography, there were bi-lateral symmetrical alveolar opacities located centrally inthe mid and lower lung zones. High resolution CT (HRCT)scanning revealed widespread ground-glass opacificationin nine cases and “crazy-paving” in the remainder. The di-agnoses were established in all patients by transbronchialbiopsy or from fluid obtained at bronchoalveolar lavage



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(BAL). A total of 29 episodes of whole lung lavage wereperformed across 12 cases, using between 15 and 30 litresper episode. Eleven cases had sequential right and left lunglavages, twice in one case. The twelth case required fiveseparate episodes of lavage on the left over a period of sixyears for progressive disease. In this case, the fourth lava-ge resulted in severe hypoxemia and prolonged intensivecare unit (ICU) stay, probably due to an excess amount ofresidue left in the lung: the fifth lavage was fatal after pro-longed ICU stay due to ventilator-associated pneumonia(VAP) and sepsis. Two of the patients experienced spon-taneous remission.

C06Acute fibrinous and organizingpneumonia (AFOP) in the ICU-casereportEduardo Sousa1, João Pedro Baptista1, Nuno Devesa1,

Lina Carvalho2, Jorge Pimentel1

1 Intensive Care Unit;2 Pathology DepartmentUniversity Hospital – [email protected]

AbstractAcute fibrinous and organizing pneumonia (AFOP) hasbeen recognized as a distinctive histologic pattern, whichdoes not meet the classic histologic criteria for diffusealveolar damage (DAD), BOOP, or eosinophilic pneu-monia (EP) and may represent an underreported variant.AFOP differs from the classic patterns of DAD and or-ganizing pneumonia in that organizing intra-alveolar fi-brin constitutes the dominant histologic finding, and di-ffers from the pattern of EP by the lack of prominenteosinophils.The authors report the case of a 35-years old man admittedto the hospital complaining of sore throat, fever, cough andmuco-purulent sputum. In the same day he was admitted tothe ICU with the hypothesis of community-acquired pneu-monia. The patient suffered head trauma a few years agoand there was no history of drug addiction, was a non-smoker and without other epidemiological risk factors. Des-pite the use of appropriate antibiotherapy and steroids, hiscondition didn’t improve. He was always kept under mecha-nical ventilation and developed multiorgan failure. Blood,tracheo-bronchial aspirate, bronchoalveolar lavage (BAL) andurine microbiological tests were negative. BAL showed ex-treme neutrophilic alveolitis (N: 75%; L: 12%; E: 2%; M:9%; CD4/CD8: 1,3) with normal cell count (133/mm3).Autoimmunity tests were negative. Lung biopsy was com-patible with AFOP.

The evolution of this patient was fatal after 20 days in theICU. According to some literature there are no identifiableclinical or histologic parameters predictive of patient ou-tcome with the exception of those who need mechanicalventilation that have a worse prognosis. An optimal therapyis not clearly identified.

C07A 45-year-old man with a persistentcoughJ Cemlyn-Jones, F Gamboa, C Robalo Cordeiro,

MF Baganha

Department of Pneumology and Allergology, University Hos-pital [email protected]

AbstractA 45-year-old man was referred to a pulmonologist becauseof a five-month history of cough productive of sputum,exertional dyspnea, wheezing and fatigue. He had no fever,chills or nocturnal sweats.The patient was a businessman. He did not smoke or abusealcohol and took no customary medication. He bred chi-ckens, pigeons, turtle-doves and canaries.The Chest radiograph revealed a bilateral reticulonodu-lar pattern and high resolution CT scan findings inclu-ded ground-glass attenuation. Lung function tests sho-wed restrictive ventilatory defect and reduced transferfactor. Bronchoalveolar lavage had significant increasein the total cell count (740.0/mm3) and contained highabsolute and relative numbers of lymphocytes (80%).Analysis of T-cell surface phenotypes revealed strongpredominance of CD 8 T cells associated with a redu-ced CD4/CD8 ratio (0.4). Laboratory findings includedprecipitating antibodies in pigeon’s feathers, droppingsand serum (titre 175).The patient was admitted to thehospital for a CT-guided needle biopsy. Histopatholo-gical evaluation of lung tissue revealed an interstitialinfiltrate of lymphocytes, plasma cells, neutrophils andeosinophils.The features of this case strongly suggest hypersensitivitypneumonitis.The culprit antigen was defined and the pigeonswere removed so as to avoid continued antigen exposure.Additionally, the patient was treated with oral methylpred-nisolone followed by gradual taper.There was significant cli-nical and functional improvement.Hypersensitivity pneumonitis is a granulomatous immuno-logic response of a sensitized host to inhaled biological ae-rosols. In the case of pigeon breeder’s disease, high levels ofbird antigen may persist for prolonged periods of time, des-pite removal of the offending birds.



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C08Diagnostic value of surgical lungbiopsy in diffuse pulmonary disease:Comparison with clinical andradiological diagnosisGonçalo F Coutinho, Rita Pancas, Eunice Magalhães,

João E Bernardo, Manuel J Antunes

Cardiothoracic Departamento de Cirurgia Cardiotorácica. Hos-pitais da Universidade de Coimbra. [email protected]

AbstractObjective: Surgical lung biopsy (SLB) is considered the fi-nal method of diagnostic modality in patients with undiag-nosed diffuse pulmonary disease. Nevertheless, the effectof SLB on the diagnosis, treatment and outcome of thepatient still remains controversial. The goal of this studywas to verify the concordance of the preliminary diagnosis,based on noninvasive or minimally invasive diagnosticmethods (HRCT scan, PET, bronchoscopy), with the pa-thological result of the SLB and the impact of the latter onthe treatment of those patients.Methods: Retrospective review of the experiences of SLBin our Institution, through VATS or mini-thoracotomy, in280 consecutive patients during the past 5 years. The initialdiagnosis was based on clinical settings, images and non in-vasive or minimally invasive diagnostic procedures and wascompared with the pathological findings. We considered 3major groups: diagnostic concordance (DC); new diagnosis(ND, including cases where there wasn’t a prior diagnosis);and inconclusive (IC).Results: Initial diagnosis included interstitial lung disease(34.6%), metastasis (33.9%), primitive neoplasm (10%), in-fectious pneumopathy (5%), others (2.5%) and no diagno-sis (14%). The pathologic diagnosis after SLB comprisedinterstitial lung disease (40.7%), metastasis (20.7%), primiti-ve neoplasm (14%), infectious pneumopathy (5.3%) andother lesions (13.6%). In 53.9% of patients there was a DC,in 40.4% a ND and 5.7% were IC. In 28.2% of the primitivepulmonary neoplasms the final diagnosis was unexpected.In 38.9% of patients with an initial metastatic diagnosis, thiswas excluded. For most patients (94.8%) with suspicion ofinterstitial disease a specific diagnosis could be made afterSLB. There wasn’t operative mortality and the morbidity was1.0%.Conclusion: SLB is a safe and accurate diagnostic toolfor pulmonary infiltrates of unknown aetiology, with mi-nimal operative morbidity and mortality. In our series,there was a significant discrepancy between the clinical/imagiologic findings and SLB, suggesting this diagnosticmodality has the “gold standard” in undiagnosed diffusepulmonary disease.

C09Constrictive bronchiolitis – case reportA Reis, A Segorbe Luís, MF Baganha

Serviço de Pneumologia/Departamento de Ciências Pneumoló-gicas e Alergológicas dos Hospitais da Universidade de [email protected]

AbstractThe authors present a 35 year-old caucasian female patient,former-smoker, who worked for 10 years in an archeologi-cal field, with dyspnea, wheezing, productive cough andmalaise in the last 5 months. Wheezing and rhonchi werepresent on chest auscultation.The complementary evaluation revealed peripheral bloodeosinophilia (9.9 % of the leucocyte differential-1000 cels/uL), mild hypoxemia (PO2-76 mmHg, breathing room air)and mild obstructive ventilatory deffect with normal diffu-sion capacity (TLCO). HRCT scan displayed diffuse groundglass attenuation, patchy peripheral opacities and air-trap-ping. Bronchoalveolar lavage showed neutrophilia (80%) andeosinophilia (8%) and a CD4/CD8 ratio of 0.9; the culturewas positive for Branhamella catarrhalis. Transbronchial lungbiopsy was inconclusive.HRCT scan after an antibiotic course, besides ground glassattenuation areas, showed pulmonary infiltrates with a di-fferent distribution.Anti-nuclear antibodies, anti-neutrophil cytoplasmic antibo-dies, immediate hypersensitivity skin test to Aspergillus, se-rum Aspergillus-specific IgE and IgG were negative.VATS lung biopsy was consistent with constrictive bronchiolitis.After two weeks of treatment with prednisone (0,75 mg/Kg/day) there was complete resolution of the symptoms aswell as normalization of ventilatory impairment.

C10Inflammatory markers in thebronchoalveolar lavage fluid andexhaled breath condensate ofpatients with pulmonary sarcoidosisJ Chorostowska-Wynimko, A Rozy, J Czerniawska,

A Stepniewska, E Puscinska, A Geremek, S Wesolowski,

D Gorecka

National Institute of Tuberculosis and Lung Diseases, Warsaw,[email protected]

AbstractThe relationship between proinflammatory markers con-centration in bronchoalveolar lavage (BAL) and expiredbreath condensate (EBC) as well as cytology of the respi-



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ratory tract obtained from BAL in sarcoidosis patients havenot been evaluated. To examine this we analysed the levelsof TNF-alpha, IL-6, VEGF, IGF-1 and PAI-1 in the EBCof patients with pulmonary sarcoidosis, also in compari-son to their concentration in BALF. The relationship be-tween cytokines concentration in EBC and BALF and otherdisease activity markers (BAL cytology, respiratory functi-on measurements) has been evaluated as well. Most cytoki-nes concentrations in breath condensate, as assessed byElisa method, were comparable to and highly correlatedwith their levels observed in BALF: respectively TNF-al-pha (3,79±1,78 pg/ml vs 3,34±2,81, r=0,7945, p<0,001),IGF-1 (7,76±5,9 ng/ml vs 6,09±4,44, r=0,9356, p<0,001),PAI-1 (0,81±0,42 pg/ml vs 0,94±0,43, r=0,8064, p<0,001).As an exception, there was no correlation between VEGFlevels in condensate (31,99±10,57 pg/ml) and BALF(27,15±.6,71, r=0,03) as well as negative correlation forIL-6 (0,226±0,08 pg/ml vs 4,08 ±0,53, r=-0,469, p<0,01).Significant positive correlation between relative macropha-ge count in BAL and condensate content of PAI-1 andTNF-alpha as well as negative with IGF-1 was observed.Pulmonary function as assessed by TLco measuremet cor-related with IGF-1 levels in condensates (r=0,725) andBALF (r=0,8215) only. Our data prove that quantitativeevaluation of inflammatory markers in the expired breathcondensate of sarcoidosis patients is possible. HoweverEBC usefulness for noninvasive monitoring of certainmarkers disease activity needs further evaluation.

C11Identyfication of t regulatory cellsin the balf by flow cytometryJoanna Domagala- Kulawik

Department of Pneumonology Warsaw Medical University. [email protected]

AbstractImmunologic alterations play important role in the patho-genesis of many lung disorders like: interstitial lung disea-ses, chronic obstructive lung disease, lung cancer. It wasshown0that regulatory T cells (Treg, CD4+/ CD25+) playimportant role in the autoimmunity and regulation of hostimmune response.The objective of this study was determination of lym-phocyte subpopulation: T regulatory cells derived frombronchoalveolar lavage fuid (BALF) and from peripheralblood (PB).Methods: We investigated patients with different lung di-seases (pulmonary fibrosis, sarcoidosis, lung cancer, leuka-emia) and healthy persons. BAL was performed routinely

and the total and differential cell count was evaluated bymicroscopic examination. Flow cytometry method with thepairs of monoclonal antibodies anti: CD14/ CD45 andanti: CD4/CD25 was used for evaluate proportion of Tregcells. Expression of Fas on Treg cells was analysed usingCD95/Cy5 antibody.Results: Treg cells were identified in the BALF and inPB, as well. The median proportion of CD4+/ CD25+cells as a percentage of lymphocytes in the BALF was6.7% (range: 0.7%- 18.6%) and in the PB was: 14% (ran-ge 3,6% – 39%). These cells composed mean 29% ofCD4+ BALF lymphocytes and 38,4% of PB CD4+ lym-phocytes. We identified two populations of CD4+/CD25+ cells: with low and high expression of CD25. Thelast may represe~t the real population of T regulatorycells. The mean proportion of CD4+/ CD25+ lympho-cytes with high intensity of fluorescence in the BALFwas 1,9% of the pool of lymphocytes and about 30% ofthe pool of CD4/CD25+ cells. About 100% of BALFTreg cells were positive to Fas receptor.Conclusion: Identification of Treg cells in the BALF is pos-sible by flow cytometry.

C1211 years of BAL experiencein RomaniaAneta Serbescu, Luminita Cervis, Cristina Teleaga

“Marius Nasta” Institute of Pneumology, BAL Laboratory, Bu-charest, [email protected]

AbstractBackground and aim of the work: Bronchoalveolar lavage(BAL) is an extremely useful method for sampling cells andmediators from lower respiratory tract. We started our rese-arch lab from zero, and built up to a national excellence cen-ter in 11 years.Method and results: 10320 products (BAL and bronchiallavage, BL) fluids have been processed, from patients agedbetween 6 and 88 years; over the 11 years of development,the bronchological sampling techniques have been impro-ved (increased vitality of up to 95-98%; recovered vol. 70-75% and recovered cells 12-40 mil), together with gainingand securing the interest and trust of the pneumologists inthe method’s capacities.Conclusions: Started in March 1995, our laboratory (theonly one with this profile in Romania) managed to dismissthe insecurities of the medical community regarding the useof BAL as a powerful diagnostic and follow-up tool; theresults will be presented following all etiological categoriesinvestigated; the technique has also been widely used for



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TB-BL, considering the still very high incidence of tubercu-losis in Romania (120 o/oooo).Outlook: The big diversity and the huge number of lungdisease cases we are faced with offers a wide perspective forresearch studies using BAL as a method for the better phy-siopathological understanding of the ensemble of eventstaking place in the human lung.

C13Humoral immune response againstmycobacterial antigens in BAL fluidfrom tuberculosis patientsU Demkow1,2, A Goljan-Geremek1, M Filewska1, B Bialas1,

TM Zielonka1, M Sobiecka1, J Ku1, E Augustynowicz-Kopec1,

Z Zwolska1, E Rowinska-Zakrzewska1

1 2Dept of Laboratory Diagnostics and Clinical Immunologyof Developmental Age, Warsaw Medical University;

2 Institute of Tuberculosis and Lung Diseases, Plocka 26, Warsaw,Poland

[email protected]

AbstractThe resistance to TB is cells-mediated but humoral responseis common and may be correlated with the lack of effectivelocal cellular defence mechanisms. The goal of the study wasto evaluate IgG, IgA and IgM mediated humoral immune res-ponse against 38kDa and 16 kDa or 38kDa and LAM myco-bacterial antigens in BAL fluid from patients with tuberculo-sis (TB) compare to non-tuberculous controls (NTB).179 BAL fluids (BALF) (56 TB, 123NTB – 74 non-specificpulmonary infections, 19 lung cancer, 14 sarcoidosis, 14mycobacterial infections other than tuberculosis, 2 healthycontrols) were examined. Commercially available ELISA –based assays were used. Cut off established according toserial dilution and ROC curves. Mean IgG level against 38 +16kDa was significantly higher in tuberculosis group com-pared to control (p<0,05 Three different dilutions of BALF:1: 1; 1: 10 and 1:50 (100) were tested. Only for dilution 1: 10obtained results allowed to differentiate TB group and con-trol group. Mean IgG level against 38 + LAM was signifi-cantly higher in TB group compared to control (p<0,0001).Mean IgA and against 38kDa + LAM level was also higherin TB group compared to NTB (p<0,05). No difference wasobserved between TB and NTB group in titer of IgM anti-bodies. The findings of the study indicate that TB is associ-ated with the presence of detectable levels of antibodies inthe BAL F. Antibody response is highly heterogeneous. Thisphenomenon results from the balance between pathogenand host immune system. Examined tests detecting IgG inBALF can be used in combination with other diagnosticmethods to increase diagnostic accuracy of pulmonary TB.

C14Immuno-diagnosis of TB using BALor induced sputumRAM Breen, SM Barry, FM Perrin, G Janossy, MCI Lipman

Royal Free and University College Medical [email protected]

AbstractObjective: To examine the role of bronchoalveolar lavage(BAL) and induced sputum (IS) in the immunological inves-tigation of possible tuberculosis (TB).Methods: BAL was incubated overnight with no antigen orPPD. Cells were fixed, permeabilised and then stained withmonoclonal antibodies against CD3, CD4 and IFN;. Thepercentage of PPD-specific IFN; synthetic CD4+ T-lym-phocytes (%CD4+IFN+) was identified using flow cyto-metry. Responses were also examined in IS and whole blood.Results: 238 individuals had BAL. 111/238 were diagno-sed with active TB with a median [range] %CD4+IFN+of 13.45% [0-79.11%]. 106/111 (95%) had %CD4+IFN+>1.5%. Median %CD4+IFN+ for 127 without active TBwas 0.10% [0-27.10%]. 97/127 (76%) had %CD4+IFN+<1.5%. Median %CD4+IFN+ in whole blood for 58 acti-ve TB cases was 0.19% [0-1.63%]. Positive and negativepredictive values for active TB using a 1.5% cut-off in BALwere 78% and 95%. The assay worked well with HIV andalso extra-pulmonary TB. In pulmonary TB assay sensiti-vity was 93% versus 77% for nucleic acid amplificationtesting. Most subjects with %CD4+IFN+ >1.5% withoutactive TB had evidence of previous TB. 10 TB patientshad paired BAL and IS. Median BAL %CD4+IFN was12.05% [2.63-61.04%] versus 11.16% [0.51-23.79%] in IS.In total, 26 TB patients had IS with median %CD4+IFN+of 4.49% [0-23.79%] (%CD4+IFN+ >0.5% in 24/26;>1%.in 22/26).Conclusions: BAL and IS can be used in a highly sensitivediagnostic TB immunoassay with advantages over blood.

C15Bronchoalveolar lavage inpulmonary tuberculosis diagnosisR Gerardo, L Flora, A Borba, A Mineiro, J Cardoso,

R Matos

Hospital de Santa [email protected]

AbstractIntroduction: Early identification and diagnosis of activepulmonary tuberculosis (PTB) cases is essential to any pro-gram intended to control the spread of this significant publichealth problem. Due to the high incidence of tuberculosis in



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Portugal (39/100 000), all the patients studied in our bron-chology unit are submited to mycobacteriological evaluation.Objectives: Evaluate the diagnostic accuracy for PTB ofbronchoalveolar lavage (BAL).Material and methods: We conducted a retrospective re-cord review of all patients with pulmonary tuberculosis di-agnosed in our bronchoscopy unit, between January 1999and December 2005, who were submited to flexible bron-choscopy with bronchial lavage (BL) and BAL. 237 patientswere included (168 males and 69 females),with mean age of44,7 years (range 20-85). 86 (36,2%) patients were HIV+.BAL and BL samples were tested for Ziehl-Niessen (ZN)and Lowenstein-Jensen (LJ).Results: 214 (90%) patients were diagnosed with BL samplesand 141 (59,5%) with BAL. 23 (10%) patients were positiveonly in BAL samples. 186 of the 214 with BL samples pre-sented LJ positivity and 28 had only ZN positive. In the 141BAL samples, 109 had positive LJ and 32 had only ZN.Conclusions: BL was more accurate in diagnosing PTB(90%) than BAL but 10% of the patients needed the BALsamples positivity for diagnosis.

C16RBILD – clinicopathologic featuresS Freitas, C Robalo Cordeiro, B Rodrigues, A Catarino,

MJ Matos, I Ferreira, L Carvalho, MF Baganha

Departamento de Ciências Pneumológicas e AlergológicasHospitais Universidade de [email protected]

AbstractBackground & aim: Respiratory Bronchiolitis (RB), des-cribed by Niewoehner et al in 1974, is a common finding inheavy smokers and is characterized by the presence of pig-mented macrophages within respiratory bronchioles andadjacent alveoli. Later, Myers et al described RB associatedwith interstitial lung disease (RBILD). The aim of this studyis to analyse a group of 8 patients with biopsy-proven RBILDdiagnosed within the last five years, assessing epidemiologi-cal data, clinical and imaging features, lung function tests,bronchoalveolar lavage findings, therapeutic approaches andclinical evolution.Results: The patients ranged from 28 to 63 yr of age andincluded 7 men and 1 woman. All were current smokers(13-60 pack years). All were symptomatic at presentation, withproductive cough and shortness of breath being the com-monest symptoms. Chest radiography showed predominan-tly reticular or reticulonodular shadowing, confirmed by CTscan but also showing ground glass opacification in 5 pati-ents. Both restrictive and obstructive patterns were found inlung function tests, with a gas transfer deficit in 4 patients.

Bronchoalveolar lavage was performed in 5 patients, revea-ling consistently a CD4/CD8 ratio lower than 1. The treat-ment of choice was smoking cessation and conservatorymeasures. All patients but 1 have remained stable or impro-ved after a mean follow-up of 22 months.Conclusions: The authors support that the diagnosis ofRBILD should require an appropriate clinical setting (inclu-ding smoking habits), specific imaging findings (like groundglass shadowing and centrilobular nodules), and the patho-logical confirmation, obligatory in the exclusion of moreadverse interstitial lung disease, namely idiopathic pulmo-nary fibrosis.

C17Differential cell count ofbronchoalveolar lavage in healthyvolunteers depends on volumeJ Müller-Quernheim, A Prasse, M Rothe, UF Senge,

H Hamm, M Ziegenhagen, G Zissel

Dep. of Pneumology, University Medical Center, Freiburg,[email protected]

AbstractBronchoalveolar lavage (BAL) is frequently used in the di-agnostic work-up of patients with suspected interstitial lungdiseases and its differential cell count may direct further cli-nical evaluation. To this end normal ranges are required.However, differential cell count depends on lavage volume,processing techniques, and smoking status. In this study wekept the BAL processing in the laboratory constant andanalyzed the influence of volume and smoking status in alarge cohort of 98 healthy individuals (57 non-smokers (NS),41 smokers (SM)). Volume was categorized in low (LO, 100to < 300 ml, median: 120 ml, n = 23), medium (ME, 300 to< 500 ml, median: 300 ml, n = 54), and large (LA, 500 to700 ml, median: 600 ml, n = 19).Recovery was similar in all volumes used ranging from 64 ± 8% in ME to 76 ± 10 % in LA. A close correlation between BALvolume and cell recovery was observed in NS and SM (p<0.0001and p<0.005 resp.) with up to 19 million and 56 million cellsper 100 ml recovered BAL of NS and SM, respectively. In bothNS and SM differential cell count was also influenced by volu-me with higher percentages of lymphocytes and lower percen-tages of alveolar macrophages in LA and vice versa in LO.Percentage of neutrophils was higher in LO than in ME andLA without significant differences between NS and SM. Onlylow numbers of eosinophils were observed without significantdifferences between volumes and smoking status.Normal ranges for the three volume categories for NS andSM will be presented.



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C18Bronchoalveolar lavage fluid (BAL)in diffuse parenchymal drug relatedlung disease: Report of a seriesM Romagnoli1, M Chilosi2, GL Casoni1, C Gurioli1,

L Serra3, V Poletti1

1 Department of Thoracic Diseases, Forlì, Italy,2 Department of Pathology, Verona, Italy,3 Department of Pathology, Forlì, [email protected]; [email protected]

AbstractBackground: Diffuse parenchymal drug related lung dise-ase is an increasing cause of potentially fatql acute or chro-nic pulmonary disease. Its identification is often complexand one of exclusion. Therefore, pathological data in corre-lation with clinical, laboratory and radiological features arehelpful and might be often all necessary together for a defi-nitive diagnosis.Subjects and Methods: We retrospectively analyzed thir-ty-four subjects (23 males, 11 females; mean age 56.8±16)with diffuse parenchymal drug related lung disease, develo-ped in 21 patients with baseline malignancy after a treat-ment with anti-neoplastic drugs in a chemotherapy regimen(oncological group), and in 13 patients with non-neoplasticbaseline disease, treated by a single agent (non-oncologicalgroup). All patients underwent fiberoptic bronchoscopy(FBS) with bronchoalveolar lavage fluid (BAL) and trans-bronchial biopsies. In 4 cases a surgical lung biopsy was alsoperformed.Results: BAL findings were represented by: an increase intotal cell number (>150.000/ml3) in 20 patients of the total34 (58.8%); lymphocytosis (lymphocytes >15% of totalcells) in 21 patients (61.7%); neutrophilia (neutrophils >4%of total cells) in 24 patients (70.5%); eosinophilia (eosino-phils >2%) in 11 patients (32.3%). A decreased CD4+/CD8+ ratio (CD4+/CD8+ <1%) was found in 20 pati-ents (58.8%), and an increased CD4+/CD8+ ratio (CD4+/CD8+ >2.4%) in 5 patients (14.7%). BAL was diagnosticper se in 8 cases of the total 34 (23.5%): 5 were lipoidpneumonia, 2 alveolar haemorrhage and 1 eosinophilicpneumonia. The most frequent histological patterns were:cellular interstitial pneumonitis (CIP) in 29.4% of cases(n=10), organizing pneumonia (OP) in 23.5% (n=8), di-ffuse alveolar damage (DAD) in 17.6% (n=6), lipoidic pneu-monia in 14.7% (n=5), the remaining being alveolar hae-morrhage, hypersensitivity pneumonitis and eosinophilicpneumonia. Patients with a DAD pattern presented signi-ficantly higher neutrophils in BAL as compared to pati-ents with CIP, OP and HP (p=0.03). Although BAL lym-phocytes showed a tendency to be higher in patients withCIP/OP/HP, they did not reach statistical significance, pro-

bably due to the low number of patients with DAD (n=6).Four oncological and 1 non-oncological patient died, allpresenting with type II reactive cells in BAL fluid, 4 with aDAD pattern, 1 with CIP.Conclusions: our series suggest that in most cases of di-ffuse parenchymal drug related lung disease, neither BALfindings nor lung biopsies are specific per se for diagnosis,but clinical, laboratory and radiological data are also neces-sary. However, BAL might be diagnostic per se in some spe-cific patterns of diffuse parenchymal drug related lung dise-ase, e.g. lipoid pneumonia, alveolar haemorrhage andeosinophilic pneumonia, without need for other invasiveprocedures. Moreover, BAL seem to have a prognostic rolein these diseases.

C19Pulmonary alveolar proteinosis(PAP): A case of successful pregnancyAna Cristina Mendes, Cliff Morgan1, M Felizardo,

Renato Sotto-Mayor, A Bugalho de Almeida

Serviço de Pneumologia Hospital Santa Maria, Lisboa. Portugal.1 Royal Brompton Hospital London [email protected]

AbstractThere are very few reports on PAP and pregnancy. We re-port a case of successful pregnancy in a patient with PAP.A 36 year old female was first symptomatic at age 19. Atage 23, after a lower respiratory tract infection, she wasdiagnosed as having PAP by pulmonary biopsy. Since then,multiple whole lung lavages (WLL) were performed whenshe was symptomatic. Otherwise she led a normal and ac-tive life despite important lung disease observed on chestx-ray and HRCT, and moderate restrictive lung disease. Aset of two consecutive WLL (right and left) were donebefore she attempted to become pregnant. By the 25th weekof pregnancy she noticed increased tiredness and she wasfound to dessaturate down to 88% with minimal exertionand she was started on supplemental oxygen with goodresult. At week 28 she was re-evaluated with HRCT whichconfirmed moderately severe PAP but no other new ordifferent pathology. By this time her SaO2 was slightlybetter and it was decided that the risks of general anaes-thesia and WLL were not justified and conservative mana-gement continued. Her SaO2 remained stable at greaterthan 90% from then on. She delivered at week 38 and 6days pregnancy. An elective caesarean section under epi-dural anaesthesia was done and a healthy baby was delive-red (Apgar 9/10). Prior to delivery her PaO2 in room airwas 52 mmHg, 24 hours after delivery was 58.4 and 3 dayslatter was 79.8 mmHg.



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C20BALF CCR5 and CXCR4 receptorsin HIV infectionM Alcide Marques1, Vera Alves, MA Santos Rosa,

MF Botelho, A Meliço-Silvestre, MF Baganha1 Pneumologic Department of Coimbra University Hospitals.

Portugal

AbstractThe discovery that specific chemokine receptors act as co-receptors for HIV, allowed a better understanding upon themechanisms underlying viral cellular entry and tropism.Aims: To evaluate the expression of CCR5 and CXCR4 inBAL fluid from HIV infected patients, submitted to bron-choscopy for detection of opportunistic lung infections.Material and Methods: 14 patients (9 males, 5 females),aged 39.6 ± 14.3 years, all HIV1 heterosexuals. In BAL fluidand in peripheral blood, we studied the receptors: CCR5,CD4 CCR5, CD8 CCR5, CCR5 M0, CXCR4, CXCR4 CD3,CXCR4 CD14, as well as the co-stimulatory molecule CD28,CD28 CD3, CD28 CD4, CD28 CD8 through monoclonalantibodies and flow citometry. In the statistical analysis, a T-Studant test and linear correlation was used.Results: Drop of CD28 co-stimulatory factor in BALf; in-crease expression of CCR5 M0 in BALf compared with theperipheral blood; statistical significant differences betweenBALf and blood for the CXCR4 (p<0.02), CXCR4 CD3(p<0.006), CXCR4 CD14 (p<0.00002).Conclusions: The lost of CD28 co-stimulatory factor is pro-bably related with immunological senescence. Both CCR5 andCXCR4 had an increased expression in BALf of these pati-ents, particularly the CCR5 M0 and CXCR4 CD14, meaninwthe important activation of lung macrophages. The over-ex-pression of these receptors may have important implicationsin future therapeutic approaches, such as, its blockage.

C21Smoking exposure effect oncytological and apoptotic parametersof BAL in smoking and NSCLCSvetlana Zunic, Snezana Zunic, Ljubisa Rakic

Clinical center of Serbia, Institute of Nuclear Medicine, Labo-ratory for Microanalytic [email protected]

AbstractSmoking exposure, as one of etiopathogenic factors of malig-nant lung diseases was investigated in relation with bronchoalve-olar lavages (BAL) cytological and apoptotic parameters. BALwas performed in13 examinees; 7 of them were patients withdiagnosis of NSCLC (Non-Small-Cell Lung Cancer). Apoptotic

parameters in BAL specimens stained by TUNEL and differen-tial cell counting of BAL specimens were performed using lightmicroscopy. Smoking exposure (pack-years) was significantly hi-gher in NSCLC patients in comparison with control smokers:44.07±21.33 vs 1.8±1.6 (p<0.001). Significant increases of eosi-nophils (p<0.01), neutrophils (p<0.05), lymphocytes (p<0.05)and decrease of alveolar macrophages (AMs) (p<0.01) were foundin patients with NSCLC in comparison with control smokers.Significant correlations of relative per cents of Foamy AMs andstage of apoptotic cells internalization by AMs and mast cells(r=-0.87, p<0.05 and r=+0.93, p<0.05, respectively) were foundin control smokers. Nonparametric correlation analysis showedthat significant correlations between Foamy AMs and apoptoticmarkers existed in control smokers. In NSCLC patients, signifi-cant correlations were found between eosinophils and mast cellsand apoptotic parameters.Thus, cumulative effect of smokingexposure might be of great interest in observation of smokinginduced tissue injury leading to oncogenesis.

C22Apoptosis of alveolar lymphocytes(AL) in interstitial lung diseases (ILD):Its extension, alterations and clinicalimportancePiotr Kopinski1, Jerzy Szczeklik3, Krzysztof Sladek2,

Barbara Balicka-Slusarczyk3, Jerzy Soja2,

Monika Swierczynska2, Grazyna Pinis4, Marta Plato1,

Monika Budna1, Karina Bartczak1

1 Chair of Gene Therapy, Collegium Medicum, Nicolaus Co-pernicus Univ, Bydgoszcz, PL

2 Chair of Internal Medicine, Collegium Medicum, JagiellonianUniv, Krakow, PL

3 Dept of Internal and Occupational Dis, Collegium Medicum,Jagiellonian Univ, Krakow, PL

4 Atopia Out-Patient Clinic, Krakow, [email protected]

AbstractBackground: Activity of inflammation in ILD depends in-ter alia on decreased apoptosis of alveolar lymphocytes, AL.The extension of this process, its relation to other inflam-matory markers, as cytokine secretion, and its clinical im-portance needs explanation.Methods: AL and AM obtained from patients with silicosis, as-bestosis, sarcoidosis (PS) and idiopathic pulmonary fibrosis (IPF)(n=13,18,57,18 resp.) were examined for: 1. cell cycle (sub-G1late apoptosis peak) 2. TUNEL 3. Annexin V, Fas, Fas Ligand(FasL), BCL-2 and caspase-3 expression. 4. BAL routine cytologyand immunotyping. BAL supernatants were tested with ELISAfor IFNã; TNFá; IL4, IGF1, soluble Fas (sFas) and soluble FasLlevels. Cell short-term cultures were performed in some cases.



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Results: Average 1% of AL enters apoptosis (and 1% prolifera-tes) in controls as detected by cell cycle analysis. Decreased ALapoptosis rate was found in Loeffgren’s syndrome (LS), progres-sive PS and asbestosis (0.6±0.2, 0,5±0.2, 0.4±0.15 vs 1.1±0.5%in controls, median ± SEM). AL apoptosis rate was negativelycorrelated with total CD4 cell number (rs -0.33, p<0.02) andCD4/CD8 ratio (rs -0.25, p<0.005). Additionally, in 3 PS pati-ents with subsequent disease remission massive AL apoptosis(up to 50% of cells) was observed. Serial BAL examinations per-formed in 7 cases of chronic PS showed increased AL apoptosistowards the rate observed in controls. Systemic steroid treatmentin 3 PS and 5 IPF patients resulted in dramatic increase of ALapoptosis rate but had no influence on AM apoptosis.TNFα levels (significantly increased in LS, active PS, IPFand asbestosis) were negatively correlated with percentage ofapoptotic AL (rs -0.31, p<0.001). Except negative correlationbetween sFas and AL apoptotic rate (p<0.05) other mediatorsseemed have no influence on BAL cell apoptosis (the observati-on proven additionally by BAL cell cultures).Conclusions: 1. AL accumulation in ILD depends in parton AL apoptosis decreased rate. 2. Inhanced AL apoptosismay be applied as a marker of clinical improvement and/orlocal impact of steroid treatment. 3. TNFα and Fas,but not IFNγ, IL4 or FasL, seems to protect BAL immunecells. 4. The major AL subset regulated in ILD lower ai-rways by apoptosis seems to be CD4 cells.

C23Expression of interferon-gamma(IFNg) in BAL fluid of patients withselected interstitial lung diseases(ILD). Better prognosis in patientswith increased CD4/CD8 ratio inpulmonary sarcoidosis (PS) andidiopathic pulmonary fibrosis (IPF)Piotr Kopinski1, Krzysztof Sladek2, Grazyna Pinis2,

Barbara Balicka-Slusarczyk3, Jerzy Soja2, Grzegorz

Przybylski4, Agnieszka Jarzemska4, Karina Bartczak1,

Joanna Golinska1 & Jerzy Szczeklik2

1 Chair of Gene Therapy, Collegium Medicum, Nicolaus Co-pernicus Univ, Bydgoszcz, PL

2 II Chair of Internal Medicine, Collegium Medicum, Jagiello-nian Univ, Krakow, PL

3 Dept of Internal and Occupational Dis, Collegium Medicum,Jagiellonian Univ, Krakow, PL

4 Dept of Chest Dis, Collegium Medicum of the Nicolaus Co-pernicus Univ, Bydgoszcz, PL

[email protected]

AbstractBackgroung: IFNγ is active as a potent antifibrotic cytoki-ne in interstitial lung diseases (ILD). T cells (acc. to Th1/

Tc1 secretion pattern) are regarded as a main local sourceof IFNγ.Methods: BAL fluid of 101 nonsmoking patients with ILD,incl. IPF(UIP), PS, extrinsic alveolitis (EA), asbestosis andsilicosis (n=17,50,7,11,16 resp.) was tested for IFNã; levels(ELISA). Results were completed with BAL immunology/cytology and referred to clinical data.Results: Increased IFNγ; levels were observed in non-trea-ted patients with EA (7.8±2.1), IPF (6.1±1.8) and PS(7.9±1.7), whereas the results of pneumoconioses were si-milar to those obtained in the controls (n=15, 2.0±1.1 pg/ml, median ± SEM). The highest IFNγ concentration wasfound in Loeffgren’s syndrome (10.2±2.4), the PS subgroupcharacterized with especially high CD4/CD8 ratio (10.5±1.2vs progressive PS 5.3±0.7; p<0.05). IFNγ level in ILD waspositively correlated with percentage of BAL lymphocyte (rSpearman +0.23; p<0.05), total CD4 cell number (rs +0.32,p<0.05) and CD4/CD8 ratio (rs +0.38, p<0.0001). In IPF/UIP group of patients with CD4/CD8<1 (n=8), IFNγ le-vel was lower as compared to the group with CD4/CD8>1(n=9): 2.8±1.3 vs 7.3±1.0 pg/ml. Steroid treatment causedinsignificant decrease in IFNã; levels of serial lavages in 7PS and 8 IPF patients.Conclusions: The results are consistent with the view onprotective role of IFNγ in ILD and may explain the betterprognosis in patients with higher CD4/CD8 ratio in ILDwith imminent fibrosis. CD4, but not CD8 seem to be themain local source of IFNγ in ILD.

C24Expression of CCX CKR in pulmonarysarcoidosisEva Kriegova, Andrey Tsyrulnyk, Arsen Arakelyan,

Frantisek Mrazek, Vitezslav Kolek, Roland M du Bois,

Helmut Popper, Martin Petrek

Palacky University Olomouc, the Czech Republic. Royal Bromp-ton Hospital London, the United Kingdom; Medical UniversityGraz, Austria;[email protected]

AbstractObjective: CCX CKR is a decoy chemokine receptor thatspecifically binds the chemokines CCL19, CCL25 and CCL21.CCL19 was previously found upregulated in pulmonary sar-coidosis. We have, therefore, investigated CCX CKR expres-sion and its plausible role in this inflammatory disease.Methods and results: CCX CKR mRNA was semiquan-titated by RT-PCR in unseparated bronchoalveolar (BAL)cells from the sarcoid patients (S, n=29) and control, heal-thy subjects (C, n=9). CCX CKR transcripts were upregu-lated in the patients (mean±SEM); S, 0.82±0.10; C,



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0.44±0.04; p=0.01. The highest number of transcripts wasobserved in patients with the most advanced chest X-ray(CXR) stage III by contrast to the initial stage I (S-I,0.72±0.10; S-II, 0.85±0.20; S-III, 1.04±0.20, p<0.02). Im-munocytochemistry localised the CCX CKR protein to ci-liated bronchial cells. Flow cytometric fluorescent liganduptake assay showed that these cells are able to internalizeits ligand.

Conclusion: The CCX CKR expression by ciliated bron-chial cells from sarcoid patients implicates this receptor inthe modulation of immune and inflammatory reactions on-going in the lung. Further investigations are required to cha-racterise the regulation and functional significance of CCXCKR expression, mechanism of internalisation and fate ofthe ligand.Grant support: GACR 310/05/2614, IGA MZ CR NR/9037.

Abstracts of Posters (P)

P01Sarcoidosis – an unusual neurologicalinvolvementE Magalhães, F Fradinho, A Franco, S Luís, MF Baganha

Departamento de Ciências Pneumológicas e Alergológicas dosHospitais da Universidade de Coimbra. [email protected]

AbstractThe authors present a 39 year-old caucasian male patient,mason, non-smoker, previously healthy until October/2002when he had two sequential episodes of uveitis respondingto topic and systemic corticosteroids. After slow taperinghe developed a neurological syndrome with nystagmus, ri-ght-sided Horner’s syndrome, hypoesthesia and ataxic he-miparesis – Wallenberg Syndrome.Cerebrospinal fluid revealed elevated protein levels. The MRIscan of the brain displayed a non-gadolinium enhanced ri-ght posterior laterobulbar lesion. The thoracic CT scanningrevealed mediastinal and hilar adenopathies. Pulmonary func-tion tests including carbon monoxide diffusion capacity(DLCO), flexible bronchoscopy and cardiac evaluation werenormal. Bronchoalveolar lavage was inconclusive. Sarcoidosiswas confirmed by mediastinal lymph node biopsy.High-dose corticosteroids were re-introduced with gradualrecovery of neurological deficits.Therapeutic tapering resulted in recurrence of ophthalmo-logic symptoms and appearance of new pulmonary lesionswith the pulmonary function tests displaying a restrictiveimpairment and reduced DLCO (50,1%).We present this case due to the exuberant neurologicalsyndrome in a patient already under systemic steroid the-rapy, discussing the therapeutic options in this type ofinvolvement.Key-Words: sarcoidosis; neurological involvement; therapeutic.

P02Is it sarcoidosis?A Campos, J Jones, P Mendes, F Gamboa, A Marques,

A Ferreira, C Leite, C Robalo Cordeiro, M Fontes Baganha

Department of Pulmonology and Alergology. Coimbra Uni-versity Hospital. [email protected]

AbstractThe authors describe a clinical case of a previously healthy39-year-old woman, with complaints of dyspnea, dry cou-gh, chest pain, fatigue and anorexia which started a monthbefore admission.The patient was a dressmaker and did not smoke.Chest radiograph showed a bilateral reticularnodular pat-tern and therefore additional work-up was carried out. SACE,serum clacium, 24 h urine calcium, pulmonary function tes-ts, DLco and 67 Ga scan were normal.Thoracic CT scan showed “... multiple well defined centi-metric nodules with precise borders spread throughout bothlung fields...”.Fibreoptic bronchoscopy showed no endoscopic abnor-malities. Cytological and microbiological analysis werenormal.Bronchoalveolar lavage (BAL) revealed: 69% mo-nocytes/macrophages, 3%neutrophils, 0.5% B lymphocytesand 14% T lymphocytes (68% CD4 T lymphocytes and26% CD8 T lymphocytes).The CD4 T cells were mostlyof the Th1 type.Serum T lymphocytes showed normal phenotype.CT-guided needle biopsy of a pulmonary nodule was per-formed but the result was inconclusive.The patient underwent surgical lung biopsy which allowedtissue diagnosis of sarcoidosis (stage III).Oral therapy with corticosteroids was prescribed and thepatient was discharged to the outpatient consultation.



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P03Role of bronchoalveolar lavagein diagnosis of interstitial lung diseases– case reportTeresa Câmara, M Alcide Marques, M Alice Pêgo,

MF Baganha

Pneumologic Department of Coimbra University Hospitals.Portugal([email protected]

AbstractSarcoidosis is a multisystem inflammatory disease of unk-nown etiology that predominantly affects the lungs andintrathoracic lymph nodes and is manifested by the pre-sence of noncaseating granulomas (NCGs) in affected or-gan tissues.Pneumoconiosis is the general term for lung disease causedby inhalation of mineral dust. Silicosis is a fibronodular lungdisease caused by inhalation of dust containing crystallinesilica (alpha-quartz or silicon dioxide), which is distributedwidely, or its polymorphs (tridymite or cristobalite), whichare distributed less widely.Bronchoalveolar lavage (BAL) is a minimally invasive diag-nostic technique that yields insights into immunologic, in-flammatory, and infeccious processes occurring at the al-veolar level being useful in the setting of interstitial lungdiseases.The authors describe the case of a 68 years old woman,which presents with insidious non-productive cough, calci-fied and non-calcified limph nodes and parenchimal conso-lidations and nodules in both lungs on CT scans. She had anhistory of occupational dust (silica) exposure.BFO with BAL and semi-distal biopsies were performed,and a diagnosis of sarcoidosis was achieved.This case emphasises the contribution of BAL in the diag-nosis of Interstitial Lung Diseases, as well as, the uncom-mon association of two fibrotic lung diseases.

P04Intersticial pneumonia associatedwith rapaycin therapy – case reportM Tátá1, L Mota1, J Reimão Pinto2

1 Hospital Pulido Valente, Lisboa, Portugal2 Hospital Curry Cabral, Lisboa, [email protected]

AbstractRapamycin is a recent immunosuppressive agent used withincreasing frequency in solid organ transplantation. Ho-wever, it has been associated with rare problems of pul-monary toxicity, such as interstitial pneumonia. These ca-

ses improved after dose-reduction or discontinuation ofthe drug. The authors present a case of a 58 year oldmale, engineer, non smoker, without occupational expo-sure risk.In 1999, he was submitted to renal transplantation for end-stage renal disease secondary to IgA nephropathy. He wastreated for five years with mycophenolate plus cyclosporineand prednisolone. Then the treatment was switched to ra-pamycin and prednisolone. Eight weeks after, he developedfever and dry cough.Computed tomography showed evidence of bilateral groundglass opacities, with lower lobes predominance.Lung function tests showed reduced carbon monoxide di-ffusion (DLCO-52, 4% and DLCO/VA-64,5%); withouthypoxaemia.Bronchoalveolar lavage (BAL) was negative for P. jiroveci,CMV, fungi, mycobacteria and bacteria. BAL showed pre-dominance of lymphocytes with a CD4/CD8 ratio of 2,6.Transbronchial lung biopsy was compatible with the hypo-thesis of interstitial pneumonia.After dose reduction of rapamycin there was clinical, func-tional and radiological improvement. Currently, about twoyears later, the patient remains stable, with good graft func-tion and normal lung tests.Key-words: interstitial pneumonia, rapamycin

P05Bronchoalveolar lavage featuresin sirolimus induced lung diseaseCarla Damas1, António Morais1, Carmo Palmares2,

Luís Delgado2

1 Serviço de Pneumologia – Hospital São João- Oporto2 Serviço de Imunologia – Faculdade de Medicina- Universida-

de do [email protected]

AbstractDrug induced lung diseases (DILD) are an increasinglycause of morbidity. Many drugs have been described, cau-sing several patterns of injury. Sirolimus is an immuno-suppressive agent increasingly used in renal and other solidorgan transplantation. Pulmonary toxicity has been re-cognized as a potential complication associated to thismedication. Interstitial pneumonitis and more rarely al-veolar haemorrhage have been described. BAL can be veryinformative in DILD either showing features that, althoughnot specific, are commonly seen or ruling out other diffe-rential diagnosis, such as infection or malignancy. Siruli-mus lung toxicity usually associates with a lymphocyticalveolitis, mainly of CD4+ type. The authors describe 4cases (3 men and 1 woman) between 46-71 years, trans-



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planted three years ago (1 patient) and 7 years ago (3 pa-tients). All of them were medicated with micofenolatomofetil (less than 1 g/day), prednisone (doses between 5and 10 mg/day) and sirolimus (2 mg/day in 3 patientsand 3 mg/day in 1). All patients referred fever in the ad-mission; 3 patients complained of dyspnoea and 2 hadproductive cough. Diffuse pulmonary infiltrates in HRCTscan were present in the four patients. BAL showed lym-phocytic alveolitis in 3 casses, however with a differentCD4/CD8 ratio (high-1, normal-1 and low-1); the BALwhich showed a high CD4+ proportion had also neutro-philia. The other patient BAL showed severe haemorrha-ge. Pulmonary infections were ruled out by specific BALstaining and cultures. After the drug suspension, all pati-ents experiment an improvement of the clinical picture.The BAL features described agree with the available data.However we found a more variability in CD4/CD8 ratiothat didn’t relate with different clinical or radiological pre-sentation or even with a different outcome.Key-words: sirolimus, lung toxicity, bronchoalveoar lavage

P06Lymphocytic intersticial pneumoniain a HIV-positive adult womanManuel Fonseca, Ana Cristina Mendes,

Renato Sotto-Mayor, Antonio Bugalho de Almeida

Hospital Santa Maria/Pulmonary Medicine Department, Lis-bon

[email protected]

AbstractLymphocytic Interstitial Pneumonia (LIP) is a rare lung di-sease characterized by infiltration of the interstitium andalveolar spaces by lymphocytes and plasma cells.It has a recognized association with AIDS, particularly inthe paediatric population.The case of a 36-yr-old HIV-positive woman, who deve-loped dyspnoea and chronic cough, ground-glass attenu-ation with areas of obliterative bronchiolitis on chest CTand severe functional impairment with hipoxemia at rest,is reported.Bronchoalveolar lavage was negative for infection and reve-aled lymphocytic cellularity. Thoracoscopic lung biopsy spe-cimens were consistent with LIP.She was started on Prednisolon 0,75mg/kg/d based on ide-al weight with marked improvement of her clinical conditi-on, radiological changes, pulmonary lung function tests andabnormal pulmonary gas exchange during formal exercisetesting.This case illustrates the rare association of LIP with HIVinfection in adults.

P07Lymphangioleiomyomatosis: A reviewof three casesC Valente, S André, C Santos, A Catarino, F Fradinho,

F Gamboa, M F Baganha

Departamento de Ciências Pneumológicas e Alergológicas dosHospitais da Universidade de [email protected]

AbstractObject of study: the authors review three cases for clinicaland radiological findings of lymphangioleiomyomatosis.Material: three female patients with 51 years, 42 years, and34 years.Methods: the clinical and radiological findings of each pa-tient were reviewed.Results: one patient had a two years right thoracalgia and dysp-nea. The chest X-ray showed a generalized, symmetric reticulo-nodular interstitial infiltrate, sparing the lung apices. High resolu-tion lung computed tomography (HRLCT) showed bilateraldiffuse thin-walled lung cysts with ground glass opacities throu-ghout all lobes. The other patient presented to the emergencyroom with a chylotorax after a car accident with traumatic ruptu-re of the thoracic duct. HRLCT showed multiple small non-confluent thin walled cysts, throughout all the pulmonary lobes,with varying sizes. The third patient presented with non-produc-tive cough, dyspnea to major exercise, asthenia, and weight loss.A chest X-ray demonstrated left pleural effusion. The diagnosticthoracocentesis revealed pleural liquid with biochemical criteriaof chylothorax. A posterior HRLCT showed multiple cysts withvarying sizes, thoracic duct ectasia, and a right pleural effusion.Conclusions: LAM is an uncommon interstitial lung diseasethat exclusively affects women, usually during their reproduc-tive years. Affected patients typically present with dyspnea andcough. LAM is characterized radiographically by interstitialreticular opacities that may be subtle or obvious and may pre-cede, accompany, or follow pneumothorax or chylothorax.The CT appearance of LAM is diagnostic when diffuse bila-teral thin-walled cysts surrounded by normal intervening lungparenchyma are demonstrated in young women.

P08Usual interstitial pneumonia– Case reportE Magalhães, F Fradinho, A Franco, S Luís, MF Baganha

Departamento de Ciências Pneumológicas e Alergológicas dosHospitais da Universidade de Coimbra. [email protected]

AbstractThe authors present a 50 year-old caucasian female pati-ent, farmer worker, non-smoker, with a gradual onset of



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persistent non-productive cough, progressive exertionaldyspnea, fatigue, malaise and weight loss. A history of ar-terial hypertension and Raynaud phenomenon of the ex-tremities was present. End-inspiratory crackles prevailingin the lung bases were detected on chest auscultation andperipheral edema was observed. The rheumatologic evalu-ation was inconclusive.The complementary evaluation revealed elevated erythro-cyte sedimentation rate and lactate dehydrogenase, positi-ve circulating anti-nuclear antibodies with a perinucleolarpattern. Antibodies anticentromere and anti Scl-70 werenegative.The chest radiograph revealed a bilateral reticular patternwith basal predominance and the high-resolution thoracicCT scanning exposed bilateral peripheral cystic images com-patible with pulmonary fibrosis and mediastinal adenopa-thies. Pulmonary function tests were consistent with a res-trictive impairment with reduced DLCO (49,5%). The restingarterial blood gases were normal and no dessaturation wasobserved during exercise. Bronchoalveolar lavage showedlymphocytosis (20%) with a normal neutrophil count (2%)and a CD4/CD8 ratio of 1,49.The cardiac evaluation demonstrated congestive heart failu-re, valvular fibrosis and circumferential pericardial effusion.The pulmonary artery pressure was normal.The nail fold microscopy was inconclusive. Esophageal ma-nometry displayed lower esophageal sphincter incompetenceand absent peristalsis in the distal esophagus.Surgical lung biopsy was performed revealing a UIP pattern.The patient was submitted to treatment with prednisone andazathioprine with persistent pulmonary symptoms and wor-sening of ventilatory impairment.At this point the patient still not fulfils the criteria for scle-roderma, witch remains the most reliable diagnostic hypo-thesis. Pulmonary and rheumatologic clinical surveillance ismaintained.Key-words: usual interstitial pneumonia; scleroderma

P09Pulmonary echinococcosis– A case reportBR Rodrigues, ER Magalhães, L Chieira, I Ribeiro,

AS Luís, MF Baganha

Hospitais da Universidade de [email protected]

AbstractEchinococcosis or hydatidosis is a zoonosis caused by thelarval stage of four Echinococcus species, belonging to fa-mily Taeniidae. Echinococcus granulosus is the most com-mon and cause cystic hydatid disease, endemic in Portugal.

Humans are one of the intermediate hosts and the mostfrequently involved organs are liver and lungs.The authors present a case report of a 76 years old patient,farmer, presented with a month-history of fever, persistentcough with cleary spumtum and a right-sided chest pain.Despite antibiotherapy there was no clinical improvement.The etiological study excluded neoplasy, tuberculosis or otherbacterial infection and confirmed the diagnosis of hepato-pulmonary cystic hydatidosis. The clinical features presen-ted in this case were strongly suggestive of a pulmonarycyst rupture.The therapeutic decision consisted in medical treatment withalbendazole, followed by surgical excision of the hydatidcysts. The hepatic cyst surgery was performed before pul-monary intervention, but the patient succumbed during theprocedure.The authors want to emphasise a clinical entity that is oftenfatal, but sometimes under appreciated.

P10Diagnostic difficulties in clinicalevaluation of interstitial lung diseases(ILD) – Two case reportsC Ch Loureiro, A Campos, A Aragão, L Carvalho,

MA Marques, MF Baganha

Coimbra University [email protected]

AbstractIntroduction: Interstitial lung pathology definite diagnosis nee-ds a transbronchic or chirurgical biopsy for histological confir-mation. Nevertheless, some clinical situations make these proce-dures difficult, namely by the clinical status of the patient or evenby their refuse. In such cases, the diagnosis is made by conside-ring clinical, imagiological and cytological information, as long asfitting in an evolutive and longitudinal classification pattern.Objective: to present the difficulties in obtaining a definiteclinical diagnosis in two case reports of ILD, the diagnosticvalue of therapeutics with positive clinical response and theprognostic value of bronchoalveolar lavage (BAL).Methods: we illustrate our objective with two case reportsof ILD:

– clinical data and test results of a 54 years old caucasianman, presenting with 8 days of pleuritic chest pain, lowfever and dry cough, after returning from a tropical coun-try and having done empirical therapeutics with azytro-micine and NSAI’s. On admission he presented diffusepulmonary opacities on chest X-Ray.

– clinical data and test results of a 52 years old caucasianman, presenting with anorexia, adynamia, myalgias, swe-



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ating and non productive cough for 5 days. While on emer-gency department he developed an ARDS needing inva-sive mechanical ventilation with a chest X-Ray showing,bilaterally, areas of parenchyma infiltration.

Conclusion: we emphasize the uncommon and severe pre-sentation of these two cases of ILD and the diagnostic andtherapeutic option we have made to achieve a clinical resolution.

P11Pulmonary alveolar proteinosisRicardo Lima, Daniela Ferreira, Ana Rego, Ana Oliveira,

Teresa Schiang, Sofia Neves

Vila Nova de Gaia Hospital Center, [email protected]

AbstractPulmonary alveolar proteinosis is a rare disorder in whichlipoproteinaceous material accumulates within alveoli. Theclinical course of the disease is variable, ranging from respi-ratory failure to spontaneous resolution. An important fea-ture of the disease is susceptibility to pulmonary infections,sometimes with opportunistic organisms.The authors present the case of a woman, 59 years-old, non-smoker sent to Pneumology consultation due to dyspnoea anddry cough. At physical examination, she had inspiratory cracklesin the inferior two-thirds of both hemithorax. The chest radio-graph had a bilateral alveolo-interstitial pattern. Computed to-mography scanning showed bilateral pulmonary consolidationand thickening of the interlobular septa, some regions of geo-graphic ground-glass. Pulmonary function tests revealed mildrestrictive defect, with reduction in carbon monoxide diffusingcapacity. Blood gases: pH-7.449; pO2-67 mmHg; pCO2- 60.5mmHg; HCO3- 24 mmol/L. The bronchoalveolar lavage hadmilky appearance, PAS-positive, phospholipoproteinaceous ma-terial suggesting alveolar proteinosis. She remained in clinical sur-veillance, without need of bronchoalveolar lavage, with sponta-neous and complete resolution after three years of follow-up.

P12Pulmonary tuberculosis presentingas a cavityC Valente, S André, A Franco, MF Baganha

Departamento de Ciências Pneumológicas e Alergológicas dosHospitais da Universidade de [email protected]

AbstractObject of study: the authors present an atypical presenta-tion of pulmonary tuberculosis.

Material: review of the clinical process of the patient.Methods: the clinical, laboratorial, and radiological findin-gs were reviewed.Results: a 43 years old male patient was referred to our depart-ment with the radiological and clinical diagnosis of right pneu-monia and partial respiratory insufficiency. The chest X-ray sho-wed a round opacity with an air-fluid level and thick walls localizedto the upper right lobe. The lung Computed Tomography de-monstrated a cavitating lesion at the posterior segment of theupper right lobe with an air fluid level. There was an associatedpneumonic infiltrate with air broncogram. During his stay hemade several antibiotics with no clinical and radiological respon-se. The examination of the sputum collection revealed no myco-bacterium tuberculosis as well as the bronchic lavage. The diag-nosis was established by the culture of this lavage that identifiedMycobacterium tuberculosis complex. The patient was referredto the local tuberculosis treatment institute.Conclusions: tuberculosis may be confused with severalother pulmonary parenchymal infections, with the same cli-nical and radiological findings. In this case the diagnosis wasmade by the culture of the bronchoalveolar lavage.

P13Tuberculosis – a cause of spontaneouspneumothoraxBG Rodrigues, ER Magalhães, L Chieira, AS Luís,

MF Baganha

Hospitais da Universidade de [email protected]

AbstractSpontaneous pneumothorax is a rare complication of acti-ve pulmonary tuberculosis, however, pulmonary fibrosis dueto tuberculous sequel is a common cause for secondary pneu-mothorax.The Authors report the case of a 27 years-old male patient,smoker, previously healthy, hospitalized with the diagnosisof right spontaneous pneumothorax with pulmonary co-llapse. Chest radiograph, performed after the placement ofa chest tube, showed expanded lungs and bilateral reticulo-nodular infiltrate.Test evaluation allowed the diagnosis of pulmonary tuber-culosis in a patient with subpleural bullae and residual fibro-tic lesions.The patient had a clinical and radiological favourable outco-me with antituberculous therapy.In this case, the authors emphasises the fact that a patientwith several risk factors for primary spontaneous pneumo-thorax – biotype, tabagism, subpleural bullae and residualfibrotic lesions, had a pneumothorax secondary to pulmo-nary tuberculosis, which is an infrequent situation.



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P14Pulmonary tb in patientwith sarcoidosisMJ Guimarães, C Alcobia, I Ribeiro, L Chieira, AS Luís,

MF Baganha

Departamento de Ciências Pneumológicas e Alergológias dosHospitais da Universidade de [email protected]

AbstractThe authors describe the clinical case of a 41 year-old man,admitted with community acquired pneumonia and hypoxe-mia in a context of classical clinical features for respiratory in-fection extended in time. Personal history for athsma control-led with appropriate medication, cutaneous sarcoidosis (oncorticotherapy for 1 year, stoped in 1999) and Perthes disease.Once the clinical diagnose of, cutaneous sarcoidosis was made,the patient went to the pulmonology physician. He ask forsome complementary study including Bronchoalveolar lava-ge who revealed a CD4+/ CD8+ of 4,5 who were determi-nant for establish the diagnosis for systemic sarcoidosis.The chest x-ray show bilateral consolidation of the lung paren-chyma. Direct microscopy was positive for AFB which condi-tioned the implementation of antituberculous agents. The spu-tum culture was positive for Mycobacterium tuberculosis.Pulmonary TB have increase incidence in immunocompromi-sed patients, and the corret diagnosis with appropriate therapyis essential in controlling the disease without lung damage.This is a clinical case of the same patient with two granulo-matous diseases.

P15pH breath condensate in atopicwheezing childrenPedro Martins1,2,3, Iolanda Caires2,3, Simões Torres4,

Joana Valente5, José Rosado Pinto1, Nuno Neuparth2,3

1 Serviço de Imunoalergologia, Hospital de Dona Estefânia2 Departamento Universitário de Fisiopatologia, Faculdade de

Ciências Médicas (UNL)3 Centro de Estudos de Patologia Respiratória (FCM, UNL)4 Serviço de Pneumologia, Hospital de São Teotónio, Viseu5 Departamento de Ambiente e Ordenamento, Universidade de

[email protected]

AbstractObjective: To compare pH in exhaled breath condensate(EBC), between atopic and non-atopic wheezing patients.Methods: 53 children with a wheezing history in the last 12months were selected at primary schools of a Portugueseinner-city (Viseu) using the ISAAC questionnaire. All of them

were submitted to eNO measurements and EBC collectionin order to evaluate bronchial inflammation. EBC pH wasanalysed on all samples after deaeration with argon. For thosewho were under asthma regular treatment, medication wasstopped three weeks before.Results: 31 males and 22 females (mean age of 7 ± 1.1years) were studied. 19 children were sensitized to at leastone airborne allergen and 34 didn’t. For the sensitized chil-dren, the median value for eNO was 22 ppb (interquartile:16-32.5 ppb) and the median value for EBC pH was 8.41(interquartile: 8.29-8.44). For the non-sensitized children, themedian value for eNO was 9 ppb (interquartile: 6.25-14.75ppb) and the median value for EBC pH was 8.40 (interquar-tile: 8.20-8.50). Statistical differences were found for eNObetween this two populations (p<0.0005). No differenceswere found for EBC pH (p: 0.65).Conclusions: EBC pH is similar on atopic and non atopicwheezing patients.Granted by: Fundação Calouste Gulbenkian, SaudAr Pro-jectSupported by: FEDER/ POCTI

P16Corticosteroids in sarcoidosis:A decision rule based approachP Lopes1, S Freitas2, J Jones2, J Figueira3, S Greco4,

C R Cordeiro2

1 Faculdade de Economia da Universidade de Coimbra2 Departamento de Ciências Pneumológicas e Alergológicas dos

HUC3 ISTL4 Facultà Economia, Universittà Catania

Hospitais Universidade de [email protected]

AbstractSarcoidosis is a multisystem disorder of unknown cause. Theclinical expression, natural history and prognosis are highlyvariable, with a tendency to wax and wane, either sponta-neously or in response to therapy, usually corticosteroids.The use of corticosteroid therapy remains controversial, al-though the perception that progressive symptomatic disea-se should be treated has become generally accepted. Thecriteria applied to evaluate this progression and monitor thedisease over time is in continuous change. Over the last ye-ars, the application of Operations Research/Decision Ai-ding techniques in Medicine has become a useful tool. Inthis paper, the authors propose a decision rule based appro-ach to derive a set of rules that will define the relevance ofa set of attributes (condition attributes), applied to identifythose patients in need of corticosteroids therapy (decision



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attribute). From a sample of 33 patients with biopsy-pro-ven Sarcoidosis, the condition attributes considered were:age, clinical features, laboratory findings, diffusion capacity,radiological stage, CT scan, bronchoalveolar lavage, Galliumscan, and extra-pulmonary involvement. Each of these at-tributes had a set of sub attributes (extra-pulmonary invol-vement assessed skin, gastrointestinal and ocular lesions (nopatients presented with CNS or cardiac involvement)). Threecondition attributes (age, radiological stage, CT scan) wereconsidered unnecessary for the decision, while the otherscorrelated in different degrees with the decision attribute.Through the application of Rough Set Theory, decision ru-les (in terms of if these symptoms/findings, then this deci-sion) were derived after computing the solution of a Boole-an function.

P17Bronchoalveolar lavagein sarcoidosis: Is there a rolefor neutrophils?Nuno Filipe Pires1, António Morais1, Cláudia Dias2,

Luís Filipe Azevedo2, Carmo Palmares1, Luís Delgado3

1 Pulmonology Department2 Biostatistics and Medical Informatics Department – Medical

School3 Immunology Department – Medical School

Hospital S. João, Porto, [email protected]

AbstractIntroduction: Lymphocytes percentage in Bronchoalveo-lar Lavage (BAL), especially CD4/CD8 ratio may be helpfulin establishing the diagnosis of sarcoidosis. Some studiessuggest a role for neutrophils, especially in what evolutionof disease is concerned. However, the real association ei-ther with sarcoidosis main characteristics or with its outco-me has not yet been established.Aim: Study of BAL neutrophils association with clinical,functional, radiological and outcome in patients with sar-coidosis.Material and Methods: 152 patients with sarcoidosis wereincluded. Clinical presentation, functional, radiological andBAL features were studied as well as the patients’ outcome.The statistical analysis was carried out by using SPSS 14.0®programme.Results: The mean age was 35 years (17-69) and predomi-nantly female (64%). Cough (48%), dyspnea (43%) anderythema nodosum (28%) were the most frequent clinicalpresentations. Lung Function Tests: normal-53%, obstruc-tive pattern-27%, restrictive pattern-13%, mixed pattern-6%.Radiological stages: I-38%, II-47%, III-8%, IV-7%. Reticu-

lonodular and micronodular were the most frequent radio-logical patterns. BAL median values: total cell count-2.6x105,lymphocytes-38%, CD4/CD8-4.5, neutrophils-1%. Extra-thoracic involvement was present in 30% of patients. A chro-nic course was observed in 45% of patients. We found sta-tistically significant differences of neutrophil percentagesamong categories of lung function tests (p=0.027), radiolo-gical stages (p=0.002) and radiological patterns (p=0.033).Significantly higher percentage were found for restrictivepattern (5.5%), radiological stages II and IV (5.8% and 5.9%respectively), reticular and fibrosis radiological patterns (5.1%and 8.2% respectively).Conclusions: According with our results neutrophils see-ms to have a role in the pathogenesis of the disease associ-ated with a more severe presentation. However consideringall the patients studied we haven’t found any influence in thenatural history of the disease.

P18The reflection of the diffusionlung capacity changes on thebronchoalveolar lavage cytologyin the newly diagnosed sarcoidosisLuminita Cervis, Cristina Teleaga, Diana Ionita,

Roxana Nemes, Nicoleta Bisca, Aneta Serbescu

“Marius Nasta” Institute of Pneumology, BAL Laboratory, SosViilor Bucharest, [email protected]

AbstractBackground: the presence of sarcoidic granulomas in thelung tissue could result in a functional impairment and inthe changes of the bronchoalveolar lavage (BAL) cellularity.The most sensitive test for the detection of the lung invol-vement is the diffusion lung capacity (DL,co).Aim: to assess whether different degrees of the diffusionlung capacity alteration are reflected in typical patterns ofBAL cytology of patients with sarcoidosis.Methods: the retrospective study included 67 patients(35males), aged 37.1±11.4 years with newly diagnosed sar-coidosis. All patients underwent chest X-ray and diffusioncapacity of the lung for carbon monoxide measurement.The patients were distributed in 4 groups depending onDL,co value: 1 with normal gas changes, 2 (DL,co 60-80%),3 (DL,co 40-60%) and 4 (DL,co < 40%). The BAL’s cellularpopulations were compared between the groups.Results: there was no statistical significant difference ofBAL’s cytological pattern between the patients with diffe-rent values of DL,co.Conclusions: the BAL’s cytological features did not reflectthe severity of the lung functional impairment.



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P19Bronchoalveolar lavagein pulmonary sarcoidosisA Rego, S Neves, A Oliveira, DS Ferreira, R Lima,

M Guimarães, A Caiado, R Duarte, S Conde, S Torres,

C Centeno, MC Brito, J Almeida, J Moura e Sá,

MC Palmares*, JL Delgado1

Serviço de Pneumologia, Centro Hospitalar Vila Nova de Gaia1 Serviço e Laboratorio de Imunologia, Faculdade de Medicina,

Universidade do [email protected]

AbstractAims: Our aim was to evaluate the demographic, clinical, radio-logical and BAL characteristics of 60 patients with histological/clinical/laboratorial evidence of pulmonary sarcoidosis who un-derwent BAL as part of their initial diagnostic evaluation. Analyzethe correlation between BAL findings and stage/prognosis.Method: Retrospective study of sarcoidosis patients’ filesthat underwent BAL. Sixty files were analyzed and the po-pulation was characterized according to age, sex, radiologi-cal staging at onset (modified Scadding criteria), DLCO, FVC,BAL results, histology confirmation and disease course. Theobtained data were analyzed using specific statistic software.Results: Mean age 36.4 yrs. Female 53.3%, male 46.7%. Radiolo-gical staging (%): stage I/II/II/IV: 35/53.3/11.7/0. BAL findings(%): lymphocytic alveolitis 75.4, normal 8.8, lymph/eosin alveolitis:5.3, lymph/neutr alveolitis 3.5, other 7.1; CD4/CD8 ratio >3.5:64.6%, increased CD8: 4.2%. DLCO mean 91.8%, FVC mean87.8%. There was histological confirmation in 51.7%. Treatmentwas started in 65%. Complete remission was achieved in 23.3%of all patients. 20% patients had progression of the disease.We did not find any relationship between neutrophylic alve-olitis and lung function or evolution of disease. Statistic sig-nificance was found between neutrophylic alveolitis andworse stage (Chi-square test, p<0.05)We did not find any relationship between CD4/CD8 ratioand stage or disease course or lung function.

P20High report of tuberculosis amongbrick workers could be anoccupational factor?Firoozeh Eslami; Majid Avijgan; Fariba Farid

Shahr-e-kord University of Medical Sciences; Islamic Republicof [email protected]

AbstractObject of Study: At present and after HIV epidemic, TB isthe second infectious disease in the world. Somebody is high

risk for TB involvement and they could be involved undersome occupational factors. One of them is Brick workerswho are exposed for some occupational and environmentalfactors which are irritant for lung. The main purpose of thisstudy is passive case finding and report of incidence andprevalence of TB among Brick workers of Isfahan Provin-ce, Iran.Materials and Method: By considering of data of centreof health department of Isfahan (2000-2005) and also dataof TB for Brick workers of Isfahan, their demographicscharacteristic have been recorded.Results: On The base of results of this study, the brickworkers are situated in 3 areas of Isfahan. In these areas thereport of incidence of TB among Brick workers are as fo-llow: Isfahan (16.2%, 33.9%), Aran-Bidgol (9.1% -20%) andBorkhar-Meymeh (14.7% -44.4%).Summaries and Conclusions: By considering of these re-sults, Borkhar-Meymeh and Aran-Bidgol are reported to havehighest (44.4%) and lowest (9.1%) of TB report.It seems this finding have a direct relation with number of bri-ckers and number of non residential workers (Afghanis) in the-se areas. On the base of these reports and also the probablerelation of TB incidence and occupational factor, it suggestshaving a more observation and following up for these workers.Key-words: Brick workers, Occupational health, TB

P21Natural killer T cells (NKT)in bronchoalveolar lavageof patients with sarcoidosisKatarina Osolnik, Peter Korošec, Izidor Kern

University Clinic of Respiratory and Allergic [email protected]

AbstractIntroduction: NKT cells are a lymphocyte subset that isdistinct from conventional T cells and NK cells. They coex-press T and NK receptors and may represent cytolytic effec-tor T cells or distinct T cell lineage such as CD1d-restrictedNKT cells, which are unique in their rapid production oflarge quantity of both Th1-biased and Th2-biased cytoki-nes, particularly IFN-, TNF and IL-4. It was recently sho-wed that NKT cells could also play an important role inimmunologically mediated interstitial lung diseases.The AIM of our study was to examine the pulmonary NKT-cellnumbers and distribution in bronchoalveolar lavage (BAL) inpatients with acute sarcoidosis manifesting as Löfgren’s syndro-me and in patients with newly diagnosed, untreated sarcoidosis.Materials and Methods: We included 62 patients (35F,27M) classified into two groups: Löfgren’s syndrome (9F,5M) and others (26F, 22M). BAL was performed with flexi-



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ble fiberoptic bronchoscope (Olympus Optical Co Ltd,Tokio, Japan). The phenotypes of BAL lymphocytes weredetermined by flow cytometric analysis in specimens with>15% of lymphocytes, using FITC or PE conjugated mo-noclonal antibodies. Natural killer T cells (CD3+CD16/56+), helper (CD3+CD4+) and suppressor (CD3+CD4+)T cells, NK cells (CD3+CD16/56+) and B lymphocytes(CD3-CD19+) were scored in percentages of lymphocytes.Results: We found slightly higher frequency of pulmonaryNKT cells in patients who were presented with Lofgren’s syn-drome (median (range); 4% (1-15)) in comparison to those withnewly diagnosed, untreated sarcoidosis (2% (0-16), but not tostatistical significance (P=0.073; Mann-Whitney test). There wasno significant difference between other lymphocytes subsets.Conclusions: These results suggest the possible role of NKTcells in regulating inflammatory process in acute sarcoidosis,however further studies are needed to confirm these results.

P22Study of medicinal effect ofroncoleikin in vitro at macrophagesof bronchoalveolar fluidEP Kalinina, NS Zhuravskaya, TP Gerasimenko

Vladivostok Branch GU of the Far Eastern Science Centre ofPhysiology and Respiration SB RAMS – The Institute of MedicalClimatology and Rehabilitation, [email protected]

AbstractIt has been worked out the model for investigation of ron-coleikin medicinal effect in vitro at phagocytes of broncho-alveolar fluid. Bronchial lavage was centrifugated at 1,5thsrevolutions per minute during 10 minutes. Roncoleikin wasadded to incubative mixture in concentration 250 and 2,5mg per 1ml of incubative mixture. Period of incubation ofbronchoalveolar lavage cellular elements with preparationmade 30 minutes at 37˚C. Phagocytic activity and res-piratory metabolism of imunnocompetent cells were esti-mated by the modified method Park, prodigiosan was usedas activator.Roncoleikin in concentration 250mg in 1 ml of incubativemixture rose phagocytic activity of immunocompetent cellsper 12,6%±0,9% (p<0,001), total quantity of actively meta-bolic phagocytes per 23%±1,4 (p<0,05) and the level ofintracellular metabolism of some cells of monocytic-ma-crophage link per 0,19%±0,04 (p<0,05). Roncoleikin addi-tion in concentration 100 times less the previous one didn’tinfluence on immunologic parameters. Roncoleikin in vitroin concentration 250mg in 1ml 0f incubative mixture im-pacts on activity and intracellular metabolism of phagocytsin bronchoalveolar fluid.

P23Comparison of alveolar cellpopulations, nk and nkt cells betweencop/boop and eosinophilic pneumoniaD Papakosta, K Manika, G Kyriazis, T Kontakiotis,

N Manolakoglou, D Gioulekas, D Bouros, D Patakas

Department of Pneumonology, Aristotle University of Thessa-loniki,G. Papanicolaou Hospital, Exohi, Thessaloniki, [email protected]

AbstractAim of the study: To evaluate bronchoalveolar lavage fluid(BALF) findings in patients with COP/BOOP and eosino-philic pneumonia (EP) and to investigate any potential di-fferences between these two groups.Patients and Methods: BALF from 59 patients (32 COP/BOOP and 27 EP, mean age 58.03 5 17.51 years)were analyzed. BALF total cells and cell differentials werecounted while CD3+, CD4+, CD8+, CD19+ and cytotoxiclymphocytes CD3-CD16/56+ (NK cells) and CD3+CD16/56+ (NKT cells) were determined by flow cytometry.CD4/CD8 ratio was also calculated.Results: Statistically significant differences between COP/BOOP and EP were observed in percentages of lympho-cytes (p= 0.000), eosinophils (p=0.000) and NKT cells(p=0.001). No differences were observed in CD4 and in CD8cells between the two groups. In patients with COP/BOOP asignificant negative correlation between NK and T- cells wasobserved (p=0.000, r=-0.684) whereas in patients with EP asignificant positive correlation between percentages of eosi-nophils and NK-cells was observed (p=0.000, r=0.846).Conclusions: Though both groups showed similar cell pro-files in CD8 and NK cytotoxic cells, they displayed differentprofiles concerning NKT cells.

P24BOOP – Experience of a pulmonology wardAna Luísa Fonseca1, Manuela Meruge2, Fernando Matos1,

Fernando Barata1, Ana Arrobas1, Yvette Martins1.1 Serviço de Pneumologia2 Serviço de Anatomia PatológicaCentro Hospitalar de Coimbra (CHC)

AbstractBOOP (Bronchiolitis Obliterans Organizing Pneumonia) is a phy-siopathologic syndrome associating suggestive clinical and ima-ging features, together with histopathologic studies showing budsof connective tissue in the lumen of the distal pulmonary airspace.Bronchoalveolar lavage (BAL) may assume characteristicpatterns and help in differential diagnose.The study is a retrospective review of all patients with BOOP,



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diagnosed in the Pulmonology Ward of CHC, between 2000and 2005.Eleven cases were diagnosed. Six patients were female and fivewere male; average age was in 54,8 years; ten patients were non-smokers and one was ex-smoker; five patients had co-morbiditi-es; mean duration of symptoms was 62,1 days; initial symptomswere dyspnea (8 patients), cough (7 patients), fever (5 patients)and weight loss (2 patients); the majority (9) of patients had beengiven multiple antibiotics before the diagnosis; crackles were heardin 6 patients, fever in 6 patients and tachypnea in 5; chest X-rayshowed bilateral alveolar opacities in 6 cases, focal pneumonia in3, linear opacities in 1 and multiple bilateral nodes in 1; lung func-tion testing, performed in 7 patients, showed a reduction in thediffusion capacity in 5 (average DLCO/VA of 67.1%); BAL wasperformed in 8 patients, and revealed an increase in the percenta-ge of lymphocytes in 7, with CD4/CD8 of 1.4 in average; histo-pathologic studies of pulmonary biopsy were diagnostic in allcases; nine patients were given corticosteroids; resolution wasfavourable in all patients except one who died.Of the parameters we looked at we emphasize: the presenceof symptoms during long periods before diagnosis and ad-ministration and failure of multiple systemic antibiotics;agreement between imaging and those published in the lite-rature; the increase in lymphocytes in the BAL, although theratio CD4/CD8 was not usefull for diagnosis; the good res-ponse to corticosteroids and non recurrence in our patients.Key-words: BOOP, Bronchoalveolar Lavage (BAL).

P25Insulin-like growth factor-I (IGF-I)expressed in lower airwaysof patients with interstitial lungdiseases (ILD) is active as mitogenof immune cells and antiapoptoticprotective agent of epitheliaPiotr Kopinski1, Jerzy Szczeklik3, Mariusz Duplaga2,Lukasz Adamek2, Barbara Balicka-Slusarczyk3,Grazyna Pinis4, Joanna Golinska1, Adam Szpechcinski1,Joanna Martenka1, Izabela Brozda1, Katarzyna Oficjalska1

Chair of Gene Therapy, Collegium Medicum, Nicolaus Coper-nicus University, Bydgoszcz, Poland1 Chair of Gene Therapy, Collegium Medicum, Nicolaus Co-

pernicus Univ, Bydgoszcz, PL2 II Chair of Internal Medicine, Collegium Medicum, Jagiello-

nian Univ, Krakow, PL3 Dept of Internal and Occupational Dis, Collegium Medicum,

Jagiellonian Univ, Krakow, PL4 Atopia Out-Patient Clinic, Krakow, [email protected]

AbstractBackground: Alveolar macrophages (AM) are known as asource of IGF-I in lower airways. Little is known about ex-

pression of IGF-I in other alveolar cells, incl. lymphocytes(AL), and about a local role of this cytokine in physiologicalconditions and in ILD.Materials and Methods: BAL was carried out in patientswith silicosis, asbestosis, idiopathic pulmonary fibrosis (IPF)and sarcoidosis (n=13, 8, 12, 56, resp). AM and AL werestudied for a) IGF-I, IGFR1, BCL-2, Fas and Fas Ligandexpression; b) cell cycle (incl. sub-G1 apoptosis peak); c)TUNEL. IGF-I levels in BAL supernatant were examinedwith ELISA. A549 cells were cultured as a model of lowerairway epithelial cells.Results: IGF-I was present in AL of all tested groups. Thenumber of IGF-I+ AL was significantly higher in IPF (52 ±6.7%) and in sarcoidosis (48 ± 5.7 vs 29 ± 6.3% in controls,p<0.05). ELISA-tested IGF-I levels were significantly in-creased in asbestosis and Loeffgren’s syndrome (18,9± 5.7and 19.1± 3.5 resp. vs 10.3± 1.3 ng/ml in controls, p<0.05).IGF-I expression in lower airways did not correlate to VCand other lung function tests. Percentage of IGF-I+ ALwas significantly correlated with parameters reflecting ALand AM cell proliferation (e.g. AL IGF-I+ vs AM in S phaseof cell cycle: rs +0.50, p=0.001) and with BCL-2 expression,but not with apoptosis. However, IGF-I antiapoptotic activitywas found in epithelial cells, since A549 cells transfectedwith vector pMT-AG-TH Anti-IGF-I (which tergets IGF-Igene promoter) demonstrated massive apoptosis, increasedpercentage of FasL+ cells and proliferation inhibition.Conclusions: 1. AL express IGF-I in normal conditionsand in ILD, especially in IPF and sarcoidosis. 2. In ourmaterial there was no evidence for profibrotic activity ofIGF-I. 3. The cytokine seems to be active as mitogen ofalveolar immune cells and antiapoptotoc protective agentfor epithelia.

P26Multinucleated giant cells in BALF– Granulomatous vs non--granulomatous ILDSilvana Smojver-Je ek1, Bo ica Vrabec-Branica1,

Tatjana Peros-Golubicic2, Jasna Tekavec-Trkanjec2

1 Department of Cytology2 Department of PulmologyUniversity Hospital for Lung Diseases “Jordanovac” Jordanovac104, Zagreb, [email protected]

AbstractAim: To estimate the frequency and type of multinucleatedgiant cells (MGC) in BAL fluid of the patients with ILDand possible differences between granulomatous and non-granulomatous diseases.



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Materials and Methods: Retrospectively 44 BAL fluid spe-cimens of patients with ILD were examined. Granuloma-tous ILD were diagnosed in 22 patients (17 sarcoidosis, 4tuberculosis and 1 histiocytosis X), and in other 22 patientsnon-granulomatous ILD (10 IPFs, 1 asbestosis, 1 eosino-philic infiltrate, 3 lung manifestations of collagen-vasculardiseases and 7 non-specified ILD). MGC were cells having3 or more nuclei, separated in groups according to cytomor-phological features and number of nuclei. Frequencies, dis-tribution of MGC in BAL fluid of granulomatous and non-granulomatous ILD and differences among frequencies inMGC groups were calculated.Results: MGC were classified in 3 groups: alveolar macro-phage–like (AM-MGC), Langhans or foreign body typeMGC (LF-MGC) and non-specific MGC (NS-MGC). Num-ber of MGC in non-granulomatous ILD was higher (n=191)than in granulomatous ILD (n=177), without statistical sig-nificance. Number of LF-MGC was statistically higher (p<0,05) in granulomatous ILD. Number of AM-MGC was sta-tistically higher (p<0,05) in non-granulomatous ILD.

P27Diversity of BAL cell profilesin interstitial lung diseasesIzidor Kern, Katarina Osolnik, Damjan Er en,

Milivoj Mermolja

University Clinic of Respiratory and Allergic Diseases, Golnik,[email protected]

AbstractCharacteristic BAL cell profiles are defined for common ILDand expressed as average values. BAL cell distribution pat-tern in ILD differs from normal type. Object of study wasto determine cell frequency distribution and to show mixedcell profiles in ILD.We analyzed 521 BAL specimens of ILD (308 sarcoidosis,60 EAA, 89 IPF, 64 CVD with lung involvment). Specimenswere representative (recovered volume of instilled fluid>30%, cell viability >60%, TCC >50 cells/μl, epithelial ce-lls <10%). Cell differentials were done by counting 200 ce-lls. Lymphocytic subpopulation were determined by flowcytometry in specimens with >15% lymphocytes. Univari-ant analysis was used to test for normal distribution, andanalysis of variance was used to compare differences betwe-en average values. Tukey’s test was used to find the meansthat are significantly different from each other.26% specimens of sarcoidosis had normal value of TCC.Lymphocytosis was observed in 76% of specimens andhalf of specimens had only lymphocytic increase whileothers were of mixed type. 45% of specimens had CD4/

CD8 >5 and 42% of specimens had normal or decreasedCD4/CD8. Neutrophils and/or eosinophils were slightlyor moderatly increased. EAA had significantly higher ave-rage value of TCC than sarcoidosis. There were more spe-cimens with the highest values of TCC. Extreme lympho-cytosis was frequently observed and was part of mixed cellprofile. Only 6% of specimens had increased CD4/CD8.Most of specimens in IPF had increased TCC. Mixed cellalveolitis was the commonest type and usually with decre-ased CD4/CD8. Similar to sarcoidosis, in CVD 25% spe-cimens had normal TCC. Lymphocytosis was observed inone third of specimens. There was mixed cell alveolitis withincrease of neutrophils, eosinophils and macrophages.CD4/CD8 was decreased.We conclude that mixed type of BAL cell profile with incre-ased TCC and decreased CD4/CD8 is the commonest inEAA, IPF and CVD. Pure lymphocytic alveolitis with signi-ficantly increased CD4/CD8 is common in sarcoidosis.

P28Pulmonary alveolar microlithiasis:12 year follow-upNesrin Mogulkoc, Serdar Soyer, Ali Veral

Pulmonary Medicine, Ege University Hospital, Izmir, TURKEY.Pathology Department, Ege University Hospital, Bornova 35100Izmir, [email protected]

AbstractPulmonary alveolar microlithiasis (PAM) is a rare idiopathiccondition that is relatively common among Turks. Abouthalf the reported cases involve more than one member of afamily, but the genetics of inheritance remains uncertain.We report on an initially asymptomatic, nonsmoking 24-yearold Turkish man, with PAM followed over a period of twel-ve years. He presented in 1994 with Guillain Barre Syndro-me, from which he made a rapid recovery. However, he wasfound to have an abnormal chest x-ray with diffuse alveolarand nodular infiltrates in both lungs. Thoracic CT showeddiffusely distributed dense micronodular opacities. Fiberopticbronchoscopy showed no endobronchial pathology. Bron-chial washings and bronchoalveolar lavage were negative foracid-fast bacilli and malignant cells but showed a microlith.A transbronchial biopsy secured a diagnosis of PAM. A ra-dioisotope bone scan showed diffuse and intense uptakethroughout both lungs. At this time, he had no respiratorysymptoms. All first-degree relatives were examined and onebrother was found to have radiologically more severe PAM.Over a period of tewelve years, the proband s FVC has de-clined an average fall of 46 ml/year. His FEV1 has fallen by70 ml/year.



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P29Suberosis: Review of clinical evolutionin two different groupsDiva Ferreira, A Oliveira, S Neves, J Almeida, JM Sá,MC Palmares1, JL Delgado1, MC Brito, S Torres1 Immunology. Hospital S. JoãoPneumology. Centro Hospitalar Vila Nova Gaia – [email protected]

AbstractBackground: Suberosis is a hypersensitivity pneumonitis-immunologically mediated, resulting from repeated inhala-tion of moldy cork dust.Aim: Characterization of two different groups in a portu-guese population with Suberosis.Material and methods: Retrospective study based on filesanalysis from patients with Suberosis at 1986 and 20 yearsafter chosen in an alleatory way. Characterization: age, sex,smoking habits, co-morbidities, clinical and radiological pre-sentation, lung funtion and bronchoalveolar lavage (BAL).Results: 32 patients, mean age of 39±4.3 yrs, male-81.2%, fema-le-18.8%. Two groups of presentation: subacute-8 (I) and chro-nic-24 (II); mean exposure of 22.4±12.2 yrs; mean time cessati-on of 10.1±3.0 yrs. We analised clinical behaviour and lungfunction at diagnosis and 20 years later. BAL and radiology wereperformed at exposure. Restrictive defect and lung diffusion(DLCO) impairment were the most frequent functional abnor-mality, particularly in group I. DLCO improved after work cessa-tion. BAL revealed high cellularity-lymphocytic alveolitis with50.64±12.7% lymphocytes. BAL CD8+ lymphocytes predomi-nated and all had CD4/CD8 ratio < 1. Lymphocitosis was hi-gher in group I – 59.56%. Ground glass opacities were the pre-dominant radiological pattern in HRCT in group I – 50%.Comments: Subacute and chronic presentations predomi-nate in Suberosis. Clinical behaviour of these groups havesubtle differences. Intensity of exposure may be important,but new biomarkers and more studies are needed.

P30Evaluation of airway inflammationby induced sputum and bronchialhyperresponsiveness in adolescentswith well-controlled asthmaAndré Moreira, Cristina Lopes, Francisco Beça,Luís Delgado, Carmo Palmares, João Fonseca,Graça Castel-BrancoImmunology Department, Biostatistics, Faculty of Medicine,University of Porto and Immunoallergology Service, H S.João,Porto, [email protected]

AbstractThis study aims to describe the correlation between indu-ced sputum inflammatory cells bronchial hyperreactivity

and lung function in adolescents with well-controlled as-thma. In 34 atopic adolescents (mean age 12.7+3.4yrs; 14females) with stable asthma, taking a median dose of 400μg inhaled corticosteroids (ICS) per day, bronchial hyper-reactivity to methacholine (PD20M), lung function (for-ced expiratory volume in one second (FEV1), and inducedsputum cellularity were assessed. Sputum was induced byhypertonic (4.5%) saline inhalation and cell counts wereexpressed as non-squamous cells percentage. Correlationsbetween variables were assessed by the Spearman correla-tion coefficient (rs). A satisfactory induced sputum sam-ple was obtained in 19 (56%) of the subjects. After spu-tum induction peak expiratory flow values were similar tothose at pre-salbutamol baseline with a difference of0.3±19.9 L/min. Sputum contained, median (range), non-squamous cells 1.0(0.2-3.3)x106ml-1, with 51.4% (10.8-69.2) macrophages, 15.5% (1.6-68.3) neutrophils, 4.0% (0.7-13.5) lymphocytes, 2.0% (0-46.1) eosinophils and 18.1%(3.1-32.4) bronchial epithelial cells. Correlations betweensputum eosinophils and PD20M (rs=-0.45, p=0.124) andFEV1 (rs=-0.40, p=0.089) were moderate but failed to re-ach statistical significance. No correlations between abso-lute cell counts and PD20M and FEV1 were observed.Sputum induction was a safe procedure although only pos-sible in a limited proportion of the subjects. In this groupof patients, with stable asthma and receiving inhaled corti-costeroids, sputum inflammatory cells were not correlatedwith bronchial hyperreactivity. These results further supportthe dissociation of bronchial inflammation and hyperre-activity in steroid treated stable allergic asthma.

P31αααααE b7 ntegrin (cd103-b7) expressionin bronchoalveolar T lymphocytesin interstitial lung diseasesAntónio Morais1, Carmo Palmares2, Luís Delgado2,João Moura e Sá3

1 Institution Pneumology Department, Hospital São João, Porto2 Immunology Department Faculdade de Medicina, Universi-

dade do Porto, Portugal3 Pneumology Department, C.H.V.N. Gaia, [email protected]

AbstractIntroduction: CD103, a receptor for the epithelial cell li-gand E-cadherin, has been reported to be preferentially ex-pressed in terminally differentiated mucosal intra-epitheliallymphocytes, and suggested to be involved in the recruit-ment of lymphocytes to lung epithelial surface. BAL CD4 Tcells expressing CD103 have been reported to be in variousamounts in diverse ILD, suggesting a role in some of thisdiseases.



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Aim: comparison of BAL CD103 expression in bronchoal-veolar T cell subpopulations in Sarcoidosis and other inters-titial lung diseases (ILD) patients.Material and Methods: CD 103 expression was investiga-ted in BAL lymphocytes of 42 patients (median age 39±16years; 20 men and 22 women). CD2, CD3, CD4, CD8, CD19and CD56 expression was also assessed by flow cytometry.Twenty one patients had Sarcoidosis with thoracic involve-ment (stage I-10, stage II- 7, stage III-2, stage IV-2). Pneu-moconiosis, lung involvement by Connective Tissue Disea-ses, Hypersensitivity Pneumonitis and Idiopathic PulmonaryFibrosis were the pathologies presented in the other 21 pa-tients. Appropriate descriptive statistics were calculated; com-parisons between groups were performed using non-para-metric Mann-Whitney U or Kruskal-Wallis tests asappropriate.Results: Sarcoidosis patients have the followed results:lymphocytes 43.2±16.4%, CD4 70.8±16, CD8

19.7±15.2, CD4/CD8 6.5±5.4, CD4+103+ 30.9±32.9(median 11.6) and CD8+103+ 43.9±32.2. Otherwise theother ILD patients revealed lymphocytes 52±24.4, CD441.2±21.9, CD8 47.7±20.6, CD4/CD8-1.3±1.3,CD4+103+ 46.7±28.8 (median-33.8) and CD8+103+66.9±27.9. We found a significant lower number of theBAL CD103+ CD4+ subset in Sarcoidosis patients,comparatively with other ILD patients (p=0.026); incontrast no statistical difference was noted in theCD8+103+ subsets.Conclusions: a selective decrease of CD103 expression cha-racterizes BAL CD4+ lymphocytes in Sarcoidosis, compa-ratively to other ILD. As CD103 is a characteristic moleculeof mucosal T lymphocytes, our data further support thehypothesis of a redistribution and alveolar compartmentali-zation of peripheral blood CD4+ lymphocytes in Sarcoido-sis, rather than an expansion of intraepithelial terminal di-fferentiated cells.



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Title Presenting Author PageC01 – Identification of bio-markers in sarcoidosis BAL using multidimensional Otto F Villa S63

Differential Display Proteome Analysis coupled with nano-HPLC MS/MSC02 – The clinical significance of IL2, IL12, TGFbeta serum and BALF levels Anna Geremek S63

in sarcoidosis. A preliminary studyC03 – Cytokine release from alveolar macrophages in idiopathic interstitial Qiao Ye S64

pneumoniaC04 – Characteristics of myofibroblasts (MyFb) recovered by Bronchoalveolar E Fireman S64

Lavage in Interstitial Lung DiseasesC05 – Pulmonary alveolar proteinosis. A report of Turkish cases Nesrin Mogulkoc S64C06 – Acute Fibrinous and organizing pneumonia (AFOP) in the ICU Eduardo Sousa S65

– Case reportC07 – A 45-year-old man with a persistent cough Jessica Jones S65C08 – Diagnostic value of surgical lung biopsy in diffuse pulmonary disease: Gonçalo Coutinho S66

Comparison with clinical and radiological diagnosisC09 – Constrictive bronchiolitis – Case report António Reis S66C10 – Inflammatory markers in the bronchoalveolar lavage fluid and Joanna Chorostowska S66

exhaled breath condensate of patients with pulmonary sarcoidosisC11 – Identification of T Regulatory Cells in the BALF by flow citometry Joanna Domagala-Kulawik S67C12 – 11 years of BAL experience in Romania Aneta Serbescu S67C13 – Humoral immune response against mycobacterial antigens in BAL Urszula Demkow S68

fluid from tuberculosis patientsC14 – Immuno-diagnosis of TB using BAL or induced sputum Ronan Breen S68C15 – Bronchoalveolar lavage in pulmonary tuberculosis diagnosis Rita Gerardo S68C16 – RBILD – Clinicopathologic features Sara Freitas S69C17 – Differential Cell Count of bronchoalveolar lavage in healthy Müller-Quernheim S69

volunteers depends on volumeC18 – Bronchoalveolar lavage fluid (BAL) in diffuse parenchymal drug Venerino Poletti S70

related lung disease: Report of a seriesC19 – Pulmonary alveolar proteinosis: a case of successful pregnancy Ana Cristina Mendes S70C20 – BALF CCR5 and CXCR4 receptors in HIV infection Alcide Marques S71C21 – Smoking exposure effect on cytological and apoptotic parameters Svetlana Zunic S71

of BAL in smoking and NSCLCC22 – Apoptosis of alveolar lymphocytes (AL) in interstitial lung diseases Piotr Kopinski S71

(ILD): its extension, alterations and clinical importanceC23 – Expression of interferon-gamma (IFNγ) in BAL fluid of patients with Piotr Kopinski S72

selected interstitial lung diseases (ILD). Better prognosis in patientswith increased CD4/CD8 ratio in pulmonary sarcoidosis (PS)and idiopathic pulmonary fibrosis (IPF)

C24 – Expression of CCX CKR in Pulmonary Sarcoidosis Eva Kriegova S72

10th International Conference on Brochoalveolar Lavage / Index: Oral Communications



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Title Presenting Author PageP01 – Sarcoidosis – An Unusual Neurological Involvement Eunice Magalhães S73P02 – Is it Sarcoidosis? Ana Campos S73P03 – Role of Bronchoalveolar Lavage in Diagnosis of Interstitial Lung Maria Teresa Câmara S74

Diseases – Case reportP04 – Interstitial Pneumonia Associated with Rapamycin Therapy – Case Report Maria Manuela Tátá S74P05 – Bronchoalveolar lavage features in sirolimus induced lung disease Carla Damas S74P06 – Lymphocytic Interstitial Pneumonia in a HIV-Positive Adult woman Manuel Fonseca S75P07 – Lymphangioleiomyomatosis:A review of three cases Carla Valente S75P08 – Usual Interstitial Pneumonia – Case Report Eunice Magalhães S75P09 – Pulmonary echinococcosis – A case report Bárbara Rodrigues S76P10 – Diagnostic Difficulties in clinical evaluation of interstitial lung Cláudia Loureiro S76

diseases (ILD) – two case reportsP11 – Pulmonary Alveolar Proteinosis Ricardo Pereira de Lima S77P12 – Pulmonary Tuberculosis presenting as a cavity Carla Valente S77P13 – Tuberculosis – A cause of spontaneous pneumothorax Bárbara Rodrigues S77P14 – Pulmonary TB in patient with sarcoidosis Maria José Guimarães S78P15 – pH breath condensate in atopic wheezing children Pedro Martins S78P16 – Corticosteroids in Sarcoidosis: a decision rule based approach Sara Freitas S78P17 – Bronchoalveolar Lavage in Sarcoidosis: is there a role for Neutrophils? Nuno Filipe Pires S79P18 – The reflection of the diffusion lung capacity changes on the Luminita Cervis S79

bronchoalveolar lavage cytology in the newly diagnosed sarcoidosisP19 – Bronchoalveolar lavage in pulmonary sarcoidosis Ana Rego S80P20 – High Report of Tuberculosis among brick workers could have Firoozeh Eslami S80

an occupational factor?P21 – Natural Killer T cells (NKT) in Bronchoalveolar Lavage of patients Katarina Osolnik S80

with SarcoidosisP22 – Study of medicinal effect of roncoleikin in vitro at macrophages Elena Kalinina S81

of bronchoalveolar fluidP23 – Comparison of Alveolar cell populations, NK and NKT cells between Despina Papakosta S81

COP/BOOP and eosinophilic PneumoniaP24 – BOOP – Experience of pulmonology ward Ana Luísa Fonseca S81P25 – Insulin-like growth factor-I (IGF-I) expressed in lower airways of patients Piotr Kopinski S82

with interstitial lung diseases (ILD) is active as mitogen of immune cellsand antiapoptotic protective agent of epithelia

P26 – Multinucleated Giant Cells in BALF-Granulomatous vs Silvana Smojver-Jezek S82Non-Granulomatous ILD

P27 – Diversity of BAL cell profiles in Interstitial Lung Diseases Izidor Kern S83P28 – Pulmonary alveolar microlithiasis: 12 year follow-up Nesrin Mogulkoc S83P29 – Suberosis: Review of clinical evolution in two different groups Diva Ferreira S84P30 – Evaluation of airway inflammation by induced sputum and bronchial André Moreira S84

hyperresponsiveness in adolescents with well-controlled asthmaP31 – αE b7 integrin (cd103-b7) expression in bronchoalveolar T lymphocytes António Morais S84

in interstitial lung diseases

10th International Conference on Brochoalveolar Lavage / Index: POSTERS

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